SMART MODE:

NORMAL GENOMIC

• Rodaway A, Patient R
• Mesendoderm. an ancient germ layer?
• Cell. 2001; 105: 169-72

• Arasu R
• In vitro reactivation of Ancylostoma caninum tissue-arrested third-stage larvae by transforming growth factor-beta.
• J Parasitol. 2001; 87: 733-8
• Display abstract

• Developmental arrest in Ancylostoma caninum is associated with preparasitic, free-living third-stage (L3) larvae, as well as anthelmintic-resilient hypobiotic L3 larvae within the tissues of an infected dog. With the tissue-arrested larvae, pregnancy and, more specifically, the hormonal effects of estrogen and prolactin mediate reactivation resulting in transmammary transmission of infection to nursing puppies. Estrogen and prolactin have been shown to be critically involved in upregulation of transforming growth factor (TGF)-beta2 during pregnancy, and studies on the soil nematode Caenorhabditis elegans further implicate TGF-beta and insulin-like signaling pathways with larval arrest and reactivation. In this report, an in vitro assay was used to show that neither estrogen, prolactin, nor insulin had a direct effect on the feeding/reactivation response of tissue-arrested larvae; however, TGF-beta isoforms 1 and 2 both had significant stimulatory effects that were comparable to the effects of dog serum. The stimulatory effects of serum could be blocked by preincubation with anti-TGF-beta antibodies. Taken together, the results support the hypothesis that during pregnancy, host-derived TGF-beta can signal a parasite-encoded receptor to trigger the reactivation of tissue-arrested larvae. TGF-beta had no effect on preparasitic larvae, suggesting that different signals may be involved in reactivation of the 2 different arrested forms of A. caninum L3 larvae.

• Miyazono K, Kusanagi K, Inoue H
• Divergence and convergence of TGF-beta/BMP signaling.
• J Cell Physiol. 2001; 187: 265-76
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily includes more than 30 members which have a broad array of biological activities. TGF-beta superfamily ligands bind to type II and type I serine/threonine kinase receptors and transduce signals via Smad proteins. Receptor-regulated Smads (R-Smads) can be classified into two subclasses, i.e. those activated by activin and TGF-beta signaling pathways (AR-Smads), and those activated by bone morphogenetic protein (BMP) pathways (BR-Smads). The numbers of type II and type I receptors and Smad proteins are limited. Thus, signaling of the TGF-beta superfamily converges at the receptor and Smad levels. In the intracellular signaling pathways, Smads interact with various partner proteins and thereby exhibit a wide variety of biological activities. Moreover, signaling by Smads is modulated by various other signaling pathways allowing TGF-beta superfamily ligands to elicit diverse effects on target cells. Perturbations of the TGF-beta/BMP signaling pathways result in various clinical disorders including cancers, vascular diseases, and bone disorders. Copyright 2001 Wiley-Liss, Inc.

• McDonnell MA, Law BK, Serra R, Moses HL
• Antagonistic effects of TGFbeta1 and BMP-6 on skin keratinocyte differentiation.
• Exp Cell Res. 2001; 263: 265-73
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• Several members of the transforming growth factor beta (TGFbeta) superfamily are expressed in the developing murine epidermis. Among these are TGFbeta1, which is found in the basal layer, and bone morphogenetic protein (BMP)-6, located in the suprabasal layers. Although the role of TGFbeta in cell growth has been studied extensively, little is known about the effects of these factors on keratinocyte differentiation. This study demonstrates that BMP-6 acts to positively regulate the differentiation of primary skin keratinocytes grown in culture. In contrast, TGFbeta1 antagonizes keratinocyte differentiation blocking the upregulation of keratin markers by BMP-6. We show that the effects of BMP-6 on expression of keratin 1 (K1), a marker of differentiation, requires signaling through the Smad pathway. In addition, regulation of K1 levels by BMP-6 is modulated by the SEK signaling pathway. This suggests that regulation of keratinocyte differentiation by BMP-6 involves multiple signaling systems. Copyright 2001 Academic Press.

• Hirai H
• [Molecular mechanism of leukemogenesis]
• Rinsho Ketsueki. 2001; 42: 162-6

• Marquez RM, Singer MA, Takaesu NT, Waldrip WR, Kraytsberg Y, Newfeld SJ
• Transgenic analysis of the Smad family of TGF-beta signal transducers in Drosophila melanogaster suggests new roles and new interactions between family members.
• Genetics. 2001; 157: 1639-48
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• Smad signal transducers are required for transforming growth factor-beta-mediated developmental events in many organisms including humans. However, the roles of individual human Smad genes (hSmads) in development are largely unknown. Our hypothesis is that an hSmad performs developmental roles analogous to those of the most similar Drosophila Smad gene (dSmad). We expressed six hSmad and four dSmad transgenes in Drosophila melanogaster using the Gal4/UAS system and compared their phenotypes. Phylogenetically related human and Drosophila Smads induced similar phenotypes supporting the hypothesis. In contrast, two nearly identical hSmads generated distinct phenotypes. When expressed in wing imaginal disks, hSmad2 induced oversize wings while hSmad3 induced cell death. This observation suggests that a very small number of amino acid differences, between Smads in the same species, confer distinct developmental roles. Our observations also suggest new roles for the dSmads, Med and Dad, in dActivin signaling and potential interactions between these family members. Overall, the study demonstrates that transgenic methods in Drosophila can provide new information about non-Drosophila members of developmentally important multigene families.

• Sasaki Y, Tsujiuchi T, Murata N, Tsutsumi M, Konishi Y
• Alterations of the transforming growth factor-beta signaling pathway in hepatocellular carcinomas induced endogenously and exogenously in rats.
• Jpn J Cancer Res. 2001; 92: 16-22
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• To elucidate involvement of the transforming growth factor-beta (TGF-beta) signaling pathway in endogenous and exogenous liver carcinogenesis, we investigated mutations of TGF-beta receptor type II (TGF-betaRII), Smad2 and Smad4 genes, and expression of TGF-betaRII in hepatocellular carcinomas (HCCs) induced by a choline-deficient L-amino acid-defined (CDAA) diet and by N-nitrosodiethylamine (DEN). Male Fischer 344 rats received a CDAA diet continuously and HCCs were sampled after 75 weeks. Administration of DEN was followed by partial hepatectomy (PH), with colchicine to induce cell cycle disturbance and a selection pressure regimen, HCCs being obtained after 42 weeks. Total RNAs were extracted from individual HCCs and mutations in TGF-betaRII, Smad2 and Smad4 were investigated by reverse transcription (RT)-polymerase chain reaction (PCR)-restriction-single-strand conformation polymorphism (SSCP) analysis followed by sequencing analysis. Mutations of Smad2 were detected in 2 out of 12 HCCs (16.7%) induced by the CDAA diet, a GGT-to-GGC transition (Gly to Gly) at codon 30 and a TCT-to-GCT (Ser to Ala) transversion at codon 118, without any TGF-betaRII or Smad4 alterations. No mutations of TGF-betaRII, Smad2 and Smad4 were encountered in eleven HCCs induced by the exogenous carcinogen. Semi-quantitative RT-PCR revealed reduced expression of TGF-betaRII in 2 HCCs (16.7%) without Smad2 mutations out of 12 HCCs induced by the CDAA diet and none of 11 induced by DEN. These results suggest that the TGF-beta signaling pathway may be disturbed in endogenous liver carcinogenesis in rats.

• Bernard DJ, Chapman SC, Woodruff TK
• An emerging role for co-receptors in inhibin signal transduction.
• Mol Cell Endocrinol. 2001; 180: 55-62
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• While many transforming growth factor-beta (TGFbeta) superfamily ligands such as TGFbeta, activin, and the bone morphogenic proteins (BMPs) are critical to the control of growth, differentiation, and cell fate, inhibin has a more limited role and is primarily responsible for the regulation of one hormone from one cell-type in the anterior pituitary. Inhibin is an endocrine hormone, produced by the gonads, that inhibits follicle stimulating hormone (FSH) release from the pituitary gonadotrope. The other hormones in the superfamily do not appear to act in an endocrine fashion, but rather control cell function in a paracrine or autocrine manner. Many components of the TGFbeta/activin/BMP signal transduction pathway have been elegantly defined; however, the mechanism of inhibin action has not been completely dissected. Several cell surface proteins that associate with inhibin have been identified recently, and these molecules may provide the clues necessary to understand how inhibin regulates reproductive function.

• Chang H, Lau AL, Matzuk MM
• Studying TGF-beta superfamily signaling by knockouts and knockins.
• Mol Cell Endocrinol. 2001; 180: 39-46
• Display abstract

• The transforming growth factor beta (TGF-beta) superfamily has profound effects on many aspects of animal development. In the last decade, our laboratory and others have performed in vivo functional studies on multiple components of the TGF-beta superfamily signal transduction pathway, including upstream ligands, transmembrane receptors, receptor-associated proteins and downstream Smad proteins. We have taken gene knockout approaches to generate null alleles of the genes of interest, as well as a gene knockin approach to replace the mature region of one TGF-beta superfamily ligand with another. We found that activin betaB, expressed in the spatiotemporal pattern of activin betaA, can function as a hypomorphic allele of activin betaA and rescue the craniofacial defects and neonatal lethal phenotype of activin betaA-deficient mice. With the knockout approach, we have shown that the expression pattern of a component in the TGF-beta superfamily signal transduction cascade does not necessarily predict its in vivo function. Two liver-specific activins, activin betaC and activin betaE are dispensable for liver development, regeneration and function, whereas ubiquitously expressed Smad5 has specific roles in the development of multiple embryonic and extraembryonic tissues.

• Bernard DJ, Chapman SC, Woodruff TK
• Mechanisms of inhibin signal transduction.
• Recent Prog Horm Res. 2001; 56: 417-50
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• Inhibin was first identified as a gonadal hormone that potently inhibits pituitary follicle-stimulating hormone (FSH) synthesis and secretion. Although the notion of a nonsteroidal, gonadally derived inhibitory substance was realized in the early 1930s (McCullagh, 1932), identification of the hormone was not accomplished until more than 50 years later. At that time, inhibin was purified from bovine and porcine follicular fluid and was shown to be produced in two forms through dimeric assembly of an alpha subunit (18 kDa) and one of two closely related beta subunits (betaA and betaB, approximately 14 kDa) (Ling et al., 1985; Miyamoto et al., 1985; Rivier et al., 1985; Robertson et al., 1985). Dimers of alpha and betaA and alpha and betaB subunits form inhibin A and inhibin B, respectively. In the process of purifying inhibin, two groups also identified homo- and heterodimers of the inhibin beta subunits (Ling et al., 1986; Vale et al., 1986). These hormones, the activins, were shown to potently stimulate FSH secretion from primary pituitary cultures and are now known to play important roles in growth and development (Woodruff, 1998; Pangas and Woodruff, 2000). Inhibins and activins are considered members of the transforming growth factor-beta (TGF-beta) superfamily of growth and differentiation factors, based on a pattern of conserved cysteine residues in the alpha and beta subunits, similar to other ligands in the family. Identification of the subunit proteins led to the cloning of their cDNAs and subsequently to their chromosomal mapping in several species (Mason et al, 1985,1986; Forage et al., 1986; Mayo et al., 1986; Esch et al., 1987; Woodruff et al., 1987; Barton et al., 1989; Hiendleder et al., 2000). Three additional activin-related beta subunits (betaC and betaE in mammals and betaD in Xenopus laevis) also have been identified but do not appear to play a role in FSH regulation (Hotten et al., 1995; Oda et al., 1995; Fang et al., 1996, 1997; Loveland et al., 1996; Schmitt el al., 1996; O'Bryan et al., 2000; Lau et al., 2000). To date, only one alpha subunit has been reported. The inhibin subunits are expressed in various tissues (Meunier et al., 1988a, 1988b) but the gonads are clearly the primary source of circulating inhibins (Woodruff et al., 1996). While inhibins act in a paracrine role in some tissues (Hsueh et al., 1987), their best-understood roles are as endocrine regulators of pituitary FSH. Activins also were purified from follicular fluid but because circulating activin levels generally are low, most actions of the hormones are likely to be paracrine in nature (Woodruff, 1998). Several reviews in the past decade have clearly and thoroughly addressed the characterization and regulation of the inhibins and activins and their roles in reproductive function (Vale et al., 1988; Ying, 1988; Woodruff and Mayo, 1990; Mayo, 1994; Woodruff and Mather, 1995). In this chapter, we focus our attention on more-recent developments in inhibin research. First, we discuss differential regulation of inhibin isoforms. Specifically, we describe patterns of inhibin A and B secretion in the context of the female reproductive cycle. Second, we review molecular mechanisms of inhibin subunit regulation. Third, while inhibins are best known for their role in pituitary FSH regulation, other functions of the ligands are becoming better understood. We review the animal and human literature addressing the possible role of inhibins in gonadal cancers. While we know "what" inhibins do in various contexts, we have a very limited understanding of "how" the ligands have their effects on target cells. Recently, candidate inhibin receptor molecules have been identified (Draper et al., 1998; Hertan et al., 1999; Lewis et al., 2000; Chung et al., 2000). Next, we detail our current understanding of inhibin signal transduction. Finally, in light of the data reviewed here, we pose questions and outline future directions for inhibin research. While this review is concerned primarily with expression and function of inhibin, activin function and mechanisms of action are described where necessary to shed light on inhibin function. Several reviews of activin's role in reproductive and other processes can be found elsewhere (Woodruff, 1998; Pangas and Woodruff, 2000).

• Morita K, Shimizu M, Shibuya H, Ueno N
• A DAF-1-binding protein BRA-1 is a negative regulator of DAF-7 TGF-beta signaling.
• Proc Natl Acad Sci U S A. 2001; 98: 6284-8
• Display abstract

• We have identified homologs of a human BMP receptor-associated molecule BRAM1 in Caenorhabditis elegans. One of them, BRA-1, has been found to bind DAF-1, the type I receptor in the DAF-7 transforming growth factor-beta pathway through the conserved C-terminal region. As analyzed using a BRA-1GFP (green fluorescent protein) fusion gene product, the bra-1 gene is expressed in amphid neurons such as ASK, ASI, and ASG, where daf-1 is also expressed. A loss-of-function mutation in bra-1 exhibits robust suppression of the Daf-c phenotype caused by the DAF-7 pathway mutations. We propose that BRA-1 represents a novel class of receptor-associated molecules that negatively regulate transforming growth factor-beta pathways.

• Kim SJ
• [Molecular mechanism of inactivation of TGF-beta receptors during carcinogenesis]
• Tanpakushitsu Kakusan Koso. 2001; 46: 111-6

• Arquier N, Perrin L, Manfruelli P, Semeriva M
• The Drosophila tumor suppressor gene lethal(2)giant larvae is required for the emission of the Decapentaplegic signal.
• Development. 2001; 128: 2209-20
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• The Drosophila tumor suppressor gene lethal(2) giant larvae (lgl) encodes a cytoskeletal protein required for the change in shape and polarity acquisition of epithelial cells, and also for asymmetric division of neuroblasts. We show here that lgl participates in the emission of Decapentaplegic (Dpp), a member of the transforming growth factor beta (TGFbeta) family, in various developmental processes. During embryogenesis, lgl is required for the dpp-dependent transcriptional activation of zipper (zip), which encodes the non-muscle myosin heavy chain (NMHC), in the dorsalmost ectodermal cells - the leading edge cells. The embryonic expression of known targets of the dpp signaling pathway, such as labial or tinman was abolished or strongly reduced in lgl mutants. lgl mutant cuticles exhibited phenotypes resembling those observed in mutated partners of the dpp signaling pathway. In addition, lgl was required downstream of dpp and upstream of its receptor Thickveins (Tkv) for the dorsoventral patterning of the ectoderm. During larval development, the expression of spalt, a dpp target, was abolished in mutant wing discs, while it was restored by a constitutively activated form of Tkv (Tkv(Q253D)). Taking into account that the activation of dpp expression was unaffected in the mutant, this suggests that lgl function is not required downstream of the Dpp receptor. Finally, the function of lgl responsible for the activation of Spalt expression appeared to be required only in the cells that produce Dpp, and lgl mutant somatic clones behaved non autonomously. We therefore position the activity of lgl in the cells that produce Dpp, and not in those that respond to the Dpp signal. These results are consistent with a same role for lgl in exocytosis and secretion as that proposed for its yeast ortholog sro7/77 and lgl might function in parallel or independently of its well-documented role in the control of epithelial cell polarity.

• Hata A
• TGFbeta signaling and cancer.
• Exp Cell Res. 2001; 264: 111-6

• Holgate ST et al.
• Invited lecture: activation of the epithelial mesenchymal trophic unit in the pathogenesis of asthma.
• Int Arch Allergy Immunol. 2001; 124: 253-8
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• BACKGROUND: A recent NIH Workshop and an ERS Task Force concluded that more work was needed to understand mechanisms of severe and chronic asthma. This report describes a series of studies that identify aberrant epithelial mesenchymal signalling in the airways as an important event in maintaining inflammation and driving remodelling in response to environmental injury. METHODS: Immunohistochemistry, genotyping and functional studies conducted on cultured asthmatic cells and mucosal biopsies were used to identify biochemical pathways involved in epithelial injury and repair in asthma and their relationship to disease severity. RESULTS: Our findings suggest that the asthmatic state results from an interaction between a susceptible epithelium and Th-2-mediated inflammation to alter the communication between the epithelium and the underlying mesenchyme - the epithelial mesenchymal trophic unit - leading to disease persistence, airway remodelling and refractoriness to corticosteroid treatment. CONCLUSIONS: Asthma is more than an inflammatory disorder, but requires engagement of important signalling pathways involved in epithelial repair and tissue remodelling. These pathways involving EGFRs and TGF-betaRs provide targets against which to develop novel therapies for chronic asthma. Copyright 2001 S. Karger AG, Basel

• Fiocchi C
• TGF-beta/Smad signaling defects in inflammatory bowel disease: mechanisms and possible novel therapies for chronic inflammation.
• J Clin Invest. 2001; 108: 523-6

• Kikuchi H, Amano H, Yamada S
• Putative role of basement membrane for dentinogenesis in the mesenchyme of murine dental papillae in vitro.
• Cell Tissue Res. 2001; 303: 93-107
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• In a new culture-conditioning system of agar-coated mesenchyme of isolated incisor dental papillae, dentinogenesis has been induced adjacent to an agar substratum that functions as a foothold for cell immobilisation. To elucidate the role of the basement membrane (BM) in dentinogenesis, we have examined the way in which dentinogenesis depends upon BM components or transforming growth factor (TGF)-beta1 in this system. At the mesenchymal-epithelial junction of odontogenic organs (cut incisor tooth germs), TGF-beta1 visibly increased in the BM during incubation. In isolated dental papillae, BM components were synthesised and deposited at aligned peripheral cells of the explants, together with an increasing amount of TGF-beta1. These components were not assembled into extracellular matrix (ECM)-absorbed agar adjacent to explants, although dentinogenesis proceeded in the presence of pericellular BM components associated with TGF-beta1. When signalling via TGF-beta type II receptors was blocked, neither ECM production nor dentinogenesis was observed but explants partially detached from the agar surface, presumably as a result of the suppressed production of ECM, since attachment was retained by pre-coating explants with artificial matrices. Rescue experiments showed that TGF-beta1 regulated dentinogenesis through ECM production. With regard to BM components, inducible dentinogenesis was Arg-Gly-Asp (RGD)-dependent. Thus, pericellular BM components associated with TGF-beta1 and an ECM-absorbed agar substratum, which affects dentinogenesis, synergistically play a role similar to that of BM components in vivo. The BM therefore serves as a structural meshwork that acts as a foothold for cell immobilisation; its components act as ligands for RGD-dependent cell adhesion and it stores TGF-beta1, which regulates ECM production.

• Kanamaru Y et al.
• Blockade of TGF-beta signaling in T cells prevents the development of experimental glomerulonephritis.
• J Immunol. 2001; 166: 2818-23
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• Anti-glomerular basement membrane (GBM) Ab-induced glomerulonephritis (GN) at late stage is thought to be mediated by T cells. However, signaling pathways of T cells that are involved in the development of anti-GBM Ab-induced GN are unclear. We have recently established transgenic mice expressing Smad7, an inhibitor of TGF-beta signaling, in mature T cells, where signaling by TGF-beta was blocked specifically in T cells. In this study, we showed that anti-GBM Ab-induced GN was suppressed in several measures in the transgenic mice including the severity of glomerular changes, proteinuria, renal function, and CD4 T cell infiltration into the glomeruli without down-regulation of CD62 ligand (CD62L) (L-selectin) expression on CD4 T cells. Furthermore, treatment with the soluble fusion protein of CD62L and IgG enhanced anti-GBM Ab-induced GN. These findings indicated that blockade of TGF-beta signaling in T cells prevented the development of anti-GBM Ab-induced GN. Because CD62L on T cells appears to be inhibitory for the development of anti-GBM Ab-induced GN, persistent expression of CD62L on CD4 T cells may explain, at least in part, the suppression of anti-GBM Ab-induced GN in the transgenic mice. Our findings suggest that the development of anti-GBM Ab-induced GN requires TGF-beta/Smad signaling in T cells.

• Hagedorn HG, Bachmeier BE, Nerlich AG
• Synthesis and degradation of basement membranes and extracellular matrix and their regulation by TGF-beta in invasive carcinomas (Review).
• Int J Oncol. 2001; 18: 669-81
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• The proper structure of the extracellular matrix, in particular of the basement membrane and the adjacent interstitial matrix, are essential prerequisites for a proper function of tissues. Invasive growth in malignant tumors is associated with a destruction of various matrix structures. Due to extensive recent analyses significant advances have been made in the knowledge of the structure of the extracellular matrix, the composition of its most important constituents, their metabolism and that of matrix degrading enzymes. This information provides insight into the pathophysiology of malignant growth. Thereby, it has been shown that malignant tumor growth is associated with a loss of basement membrane (BM) material which, however, disappears not homogeneously, but affects various BM components to different degree. The loss of an intact BM as the first barrier is therefore the initial step of tumor invasion. Despite this loss there is evidence that the de novo synthesis of BM constituents in tumor and adjacent stromal cells is enhanced. Thus, it is obvious that BM material is degraded during the invasion process to significant degree. In addition, since there is a positive correlation between the amount of retained peritumoral BM and a higher degree of tumor cell differentiation the amount of retained BM material seems to represent a marker for the biological behaviour of the tumor cells. The loss of BM material is well explained by a significant expression of major matrix degrading enzymes, the matrix metalloproteinases (MMPs) both on the mRNA and protein level. Here again, there is considerable data indicating that both tumor and stroma cells are involved in the MMP synthesis. In addition to the loss of BM substances, the interstitial extracellular matrix (ECM) is disarranged. This disarrangement may comprise enhanced de novo synthesis ("desmoplasia") or dissolution by distinct MMPs (collagenases, such as MMP-1) reflecting obviously different reaction statuses of the stromal cells. Finally, significant work has been done on the elucidation of the role of regulating cytokine systems. To this regard, particular attention has been paid to the TGF-beta system and it has been shown that the major three isoforms of TGF-betas are upregulated both in tumor and stroma cells. Since the TGF-beta-effect is mainly mediated by a particular signalling system via the TGF-beta-receptors (TBRs), the investigation of this system has provided considerable insight into the role of TBRs which are now known to represent the most potent tumor suppressor genes. Thus frequent mutations in the TBR-II gene, one of the three TBRs, in various carcinomas suggest that these molecular alterations are responsible for both the loss of the control of cellular proliferation (in tumor cells) and altered matrix metabolism (in tumor and stroma cells). The further analysis of this major cytokine system therefore will provide us with major insights into the molecular abnormalities of invasive tumor growth.

• Sturlan S et al.
• Interleukin-10-deficient mice and inflammatory bowel disease associated cancer development.
• Carcinogenesis. 2001; 22: 665-71
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• Interleukin-10-deficient mice develop colitis and colorectal cancer similar to the inflammatory bowel disease associated cancer in humans. The aim of this study was to identify possible mutations of oncogenes and tumour suppressor genes involved in tumorigenesis in Interleukin-10 (IL-10)-deficient mice. Twenty colon carcinomas from IL-10-deficient mice were screened for mutations in the K-ras and p53 genes by 'cold' single-strand-conformation polymorphism. Immunohistochemical staining was performed to detect mutations in the proteins P53, APC and MSH2, and the transforming growth factor beta type II receptor. Microsatellite instability was analysed at eight chromosomal loci and plasma levels of transforming growth factor beta1 (TGF-beta1) were also measured. At 9 weeks, 14% of the animals developed colorectal cancer, and at 10-31 weeks the incidence of carcinoma was 65%. No mutations were detected in the analysed oncogene and tumour suppressor genes. Plasma TGF-beta1 levels in IL-10-deficient mice 10-31 weeks old were higher than in wild-type littermates e.g. 45.7 +/- 4.6 ng/ml versus 19.8 +/- 4.5 ng/ml (P<0.01). No alterations in K-ras, p53, APC: and Msh2 genes suggests that other genes are involved in the development of these tumours. Elevated TGF-beta1 plasma levels correspond to the high incidence of dysplasia and cancer. Normal expression of the TGF-beta II receptors hints at genetic alterations in other members of the TGF-beta receptor signal transduction pathway.

• Eickelberg O et al.
• Molecular mechanisms of TGF-(beta) antagonism by interferon (gamma) and cyclosporine A in lung fibroblasts.
• FASEB J. 2001; 15: 797-806
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• Lung fibrosis is a fatal condition of excess extracellular matrix (ECM) deposition associated with increased transforming growth factor beta (TGF-beta) activity. Although much is known about its pathological features, our understanding of the signal transduction pathways resulting in increased ECM and collagen deposition in response to TGF-beta is still incompletely defined. We have previously reported that a JunD homodimer of the transcription factor AP-1 is specifically activated by TGF-beta in lung fibroblasts. Here we demonstrate that JunD is also specifically required for TGF-beta-induced effects. Antisense against JunD, but not c-fos or c-jun, significantly inhibited collagen deposition in response to TGF-beta in primary human lung fibroblasts. We then investigated the ability of pharmacological agents to inhibit TGF-beta-induced signaling and collagen deposition. Cs-A and IFN-gamma, but not glucocorticoids, cyclophosphamide, or azathioprine, inhibited TGF-beta-induced signaling, as assessed by luciferase reporter gene assays, and collagen deposition. TGF-beta antagonism by Cs-A was associated with direct inhibition of JunD activation, as demonstrated by electrophoretic mobility shift analyses. In contrast, the effects of IFN-gamma required signal transducer and activator of transcription (STAT)-1. We thus identify the JunD isoform of AP-1 as an essential mediator of TGF-beta-induced effects in lung fibroblasts. TGF-beta-induced signaling and collagen deposition are efficiently antagonized by Cs-A and IFN-gamma treatment, both of which exhibit distinct molecular mechanisms of action. These observations therefore offer novel targets for future therapy of fibrotic lung disease.

• Hanai J, Miyazono K
• [PEBP2/CBF is a nuclear target of Smads in TGF-beta super family signaling]
• Tanpakushitsu Kakusan Koso. 2000; 45: 48-54

• Limper AH
• Detection of TGF-beta in body fluids and tissues.
• Methods Mol Biol. 2000; 142: 39-54

• Mehra A, Attisano L, Wrana JL
• Characterization of Smad phosphorylation and Smad-receptor interaction.
• Methods Mol Biol. 2000; 142: 67-78

• Hocevar BA, Howe PH
• Regulation of AP-1 activity by TGF-beta.
• Methods Mol Biol. 2000; 142: 97-108

• Myeroff LL, Markowitz S
• Detection of TGF-beta type II receptor hot-spot mutations: the BAT-RII assay.
• Methods Mol Biol. 2000; 142: 133-7

• Massague J, Wotton D
• Transcriptional control by the TGF-beta/Smad signaling system.
• EMBO J. 2000; 19: 1745-54

• Bottner M, Krieglstein K, Unsicker K
• The transforming growth factor-betas: structure, signaling, and roles in nervous system development and functions.
• J Neurochem. 2000; 75: 2227-40
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• Transforming growth factor-betas (TGF-betas) are among the most widespread and versatile cytokines. Here, we first provide a brief overview of their molecular biology, biochemistry, and signaling. We then review distribution and functions of the three mammalian TGF-beta isoforms, beta1, beta2, and beta3, and their receptors in the developing and adult nervous system. Roles of TGF-betas in the regulation of radial glia, astroglia, oligodendroglia, and microglia are addressed. Finally, we review the current state of knowledge concerning the roles of TGF-betas in controlling neuronal performances, including the regulation of proliferation of neuronal precursors, survival/death decisions, and neuronal differentiation.

• Brown TL, Patil S, Howe PH
• Analysis of TGF-beta-inducible apoptosis.
• Methods Mol Biol. 2000; 142: 149-67

• Serrano M, Massague J
• Networks of tumor suppressors. Workshop: tumor suppressor networks.
• EMBO Rep. 2000; 1: 115-9

• Dallas SL
• Measuring interactions between ECM and TGF beta-like proteins.
• Methods Mol Biol. 2000; 139: 231-43

• Alfred J
• Development biology. Nodal signalling gets foxy.
• Nat Rev Genet. 2000; 1: 87-87

• Dong C, Li Z, Alvarez R Jr, Feng XH, Goldschmidt-Clermont PJ
• Microtubule binding to Smads may regulate TGF beta activity.
• Mol Cell. 2000; 5: 27-34
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• Smad proteins are intracellular signaling effectors of the TGF beta superfamily. We show that endogenous Smad2, 3, and 4 bind microtubules (MTs) in several cell lines. Binding of Smads to MTs does not require TGF beta stimulation. TGF beta triggers dissociation from MTs, phosphorylation, and nuclear translocation of Smad2 and 3, with consequent activation of transcription in CCL64 cells. Destabilization of the MT network by nocodazole, colchicine, or a tubulin mutant disrupts the complex between Smads and MTs and increases TGF beta-induced Smad2 phosphorylation and transcriptional response in CCL64 cells. These data demonstrate that MTs may serve as a cytoplasmic sequestering network for Smads, controlling Smad2 association with and phosphorylation by activated TGF beta receptor I, and suggest a novel mechanism for the MT network to negatively regulate TGF beta function.

• Miyazono K
• TGF-beta/SMAD signaling and its involvement in tumor progression.
• Biol Pharm Bull. 2000; 23: 1125-30
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• Cytokines of the transforming growth factor-beta (TGF-beta) superfamily are multifunctional peptides that regulate growth and differentiation of various types of cells. Members of the TGF-beta superfamily bind to type 11 and type I serine/threonine kinase receptors, which mediate intracellular signals through SMAD proteins. Of 3 subtypes of SMADs, receptor-regulated SMADs are phosphorylated by the serine/threonine kinase receptors, form complexes with common-mediator SMAD, and move into the nucleus, where they act as components of transcription factor complexes. Abnormalities of the TGF-beta receptors and SMADs have been detected in various tumors, including colorectal cancers and pancreatic cancers. Inhibitory SMADs and transcriptional co-repressors, including c-Ski and SnoN, repress the TGF-beta/SMAD signaling. Perturbation of the TGF-beta/SMAD signaling pathway may result in progression of tumors through resistance of the cells to the growth inhibition induced by TGF-beta.

• de Caestecker MP, Piek E, Roberts AB
• Role of transforming growth factor-beta signaling in cancer.
• J Natl Cancer Inst. 2000; 92: 1388-402
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• Signaling from transforming growth factor-beta (TGF-beta) through its unique transmembrane receptor serine-threonine kinases plays a complex role in carcinogenesis, having both tumor suppressor and oncogenic activities. Tumor cells often escape from the antiproliferative effects of TGF-beta by mutational inactivation or dysregulated expression of components in its signaling pathway. Decreased receptor function and altered ratios of the TGF-beta type I and type II receptors found in many tumor cells compromise the tumor suppressor activities of TGF-beta and enable its oncogenic functions. Recent identification of a family of intracellular mediators, the Smads, has provided new paradigms for understanding mechanisms of subversion of TGF-beta signaling by tumor cells. In addition, several proteins recently have been identified that can modulate the Smad-signaling pathway and may also be targets for mutation in cancer. Other pathways such as various mitogen-activated protein kinase cascades also contribute substantially to TGF-beta signaling. Understanding the interplay between these signaling cascades as well as the complex patterns of cross-talk with other signaling pathways is an important area of investigation that will ultimately contribute to understanding of the bifunctional tumor suppressor/oncogene role of TGF-beta in carcinogenesis.

• Massague J
• How cells read TGF-beta signals.
• Nat Rev Mol Cell Biol. 2000; 1: 169-78
• Display abstract

• Cell proliferation, differentiation and death are controlled by a multitude of cell-cell signals, and loss of this control has devastating consequences. Prominent among these regulatory signals is the transforming growth factor-beta (TGF-beta) family of cytokines, which can trigger a bewildering diversity of responses, depending on the genetic makeup and environment of the target cell. What are the networks of cell-specific molecules that mould the TGF-beta response to each cell's needs?

• Walker RA
• Transforming growth factor beta and its receptors: their role in breast cancer.
• Histopathology. 2000; 36: 178-80

• Lopez-Casillas F
• [Transforming growth factor-beta (TGF-beta), keystone in diabetic nephropathy]
• Rev Invest Clin. 2000; 52: 487-90

• Reinecke H et al.
• Transforming growth factor beta and coronary artery disease.
• Nutr Metab Cardiovasc Dis. 2000; 10: 101-2

• Dembic Z
• Second International Conference on Signal Transduction, Croatia May 2000.
• Scand J Immunol. 2000; 52: 641-3

• Sze JY, Victor M, Loer C, Shi Y, Ruvkun G
• Food and metabolic signalling defects in a Caenorhabditis elegans serotonin-synthesis mutant.
• Nature. 2000; 403: 560-4
• Display abstract

• The functions of serotonin have been assigned through serotonin-receptor-specific drugs and mutants; however, because a constellation of receptors remains when a single receptor subtype is inhibited, the coordinate responses to modulation of serotonin levels may be missed. Here we report the analysis of behavioural and neuroendocrine defects caused by a complete lack of serotonin signalling. Analysis of the C. elegans genome sequence showed that there is a single tryptophan hydroxylase gene (tph-1)-the key enzyme for serotonin biosynthesis. Animals bearing a tph-1 deletion mutation do not synthesize serotonin but are fully viable. The tph-1 mutant shows abnormalities in behaviour and metabolism that are normally coupled with the sensation and ingestion of food: rates of feeding and egg laying are decreased; large amounts of fat are stored; reproductive lifespan is increased; and some animals arrest at the metabolically inactive dauer stage. This metabolic dysregulation is, in part, due to downregulation of transforming growth factor-beta and insulin-like neuroendocrine signals. The action of the C. elegans serotonergic system in metabolic control is similar to mammalian serotonergic input to metabolism and obesity.

• Massague J, Chen YG
• Controlling TGF-beta signaling.
• Genes Dev. 2000; 14: 627-44

• Attisano L, Wrana JL
• Smads as transcriptional co-modulators.
• Curr Opin Cell Biol. 2000; 12: 235-43
• Display abstract

• The Smad signalling pathway is critical for transmitting transforming growth factor-beta (TGF-beta) superfamily signals from the cell surface to the nucleus. In the nucleus, Smads regulate transcriptional responses by recruiting co-activators and co-repressors to a wide array of DNA-binding partners. Thus, Smads function as transcriptional co-modulators to regulate TGFbeta-dependent gene expression.

• Stemple DL
• Vertebrate development: the fast track to nodal signalling.
• Curr Biol. 2000; 10: 8436-8436
• Display abstract

• The transcription factors Fast-1 and Mixer are important terminal components of Smad2-mediated TGF-beta signal transduction. Recent studies demonstrate that Fast-1 and Mixer play critical roles in the formation of endoderm and dorsal mesoderm in zebrafish.

• Wells RG
• Fibrogenesis. V. TGF-beta signaling pathways.
• Am J Physiol Gastrointest Liver Physiol. 2000; 279: 84550-84550
• Display abstract

• Transforming growth factor (TGF)-beta is a multifunctional peptide growth factor with a wide range of potential effects on growth, differentiation, extracellular matrix deposition, and the immune response. General TGF-beta signaling pathways have been described in detail over the last several years, but factors that determine the nature of the TGF-beta response are poorly understood. In particular, signaling pathways that specifically mediate the matrix effects of TGF-beta have received little attention, although they will be important therapeutic targets in the treatment of pathological fibrosis. This themes article focuses on TGF-beta signaling and highlights potential points for generating matrix-specific responses.

• Beppu H, Miyazono K
• [The roles of TGF-beta superfamily in mouse early development]
• Tanpakushitsu Kakusan Koso. 2000; 45: 2109-15

• Jones E, Symes A
• Leptin and TGF-beta synergistically regulate VIP cytokine response element transcription.
• Neuroreport. 2000; 11: 4049-53
• Display abstract

• Leptin was originally described as an adipocyte-derived cytokine that signals to the hypothalamus to regulate food intake and energy expenditure. Leptin signals through the Ob receptor, which is closely related to the gp130 cytokine receptor. Here we show that leptin can induce expression of the neuropeptide gene vasoactive intestinal peptide (VIP) through the VIP cytokine response element, the same element that mediates the response to the gp130 cytokines. Leptin acts synergistically with TGF-beta to activate transcription through this element. Transcriptional responses to leptin are increased when transmitted through ObR mutated at Tyr986, the SHP-2 docking domain, yet this mutation does not alter the synergy between TGF-beta and leptin. These data emphasize the functional similarity between leptin and the gp130 cytokines.

• Nishita M et al.
• Interaction between Wnt and TGF-beta signalling pathways during formation of Spemann's organizer.
• Nature. 2000; 403: 781-5
• Display abstract

• Members of the Wnt and TGF-beta superfamilies regulate both cell fate and proliferation during development and tissue maintenance. In the early amphibian embryo, the Wnt and TGF-beta superfamily signalling cascades are required for the establishment of a dorsal signalling centre, Spemann's organizer. Intracellular proteins of both pathways, upon activation, translocate to the nucleus to participate in transcription. Here we show that beta-catenin and Lef1/Tcf, which are downstream components of the Wnt signalling cascade, form a complex with Smad4, an essential mediator of signals initiated by members of the TGF-beta growth factor superfamily. In Xenopus, this interaction directly and synergistically affects expression of the twin (Xtwn) gene during formation of the organizer. This is, to our knowledge, the first demonstration of a physical interaction between TGF-beta and Wnt signalling components in vivo.

• Letterio JJ
• Murine models define the role of TGF-beta as a master regulator of immune cell function.
• Cytokine Growth Factor Rev. 2000; 11: 81-7
• Display abstract

• Many members of transforming growth factor-beta (TGF-beta) superfamily, including not only TGF-beta, but also the activins, and bone morphogenetic proteins (BMPs), have been demonstrated to affect the development and function of immune cells. From the proliferation and differentiation of pluripotent stem cells, to the activation and migration of mature lymphoid and myeloid lineages, the TGF-betas have been recognized for their ability to modulate the manner in which such cells respond to stimuli in their environment. Recent studies involving disruption of this pathway in genetically engineered mice now emphasize the importance of this activity and validate functional models predicted by in vitro studies. Phenotypic differences between mice harboring mutations in the TGF-beta1 ligand and the TGF-beta receptor-activated signaling intermediate Smad3 are presented and serve to highlight the valuable role of these in vivo genetic tests of function.

• Grainger DJ, Mosedale DE, Metcalfe JC
• TGF-beta in blood: a complex problem.
• Cytokine Growth Factor Rev. 2000; 11: 133-45
• Display abstract

• The cytokine transforming growth factor-beta (TGF-beta) was initially purified from human platelets, a rich source of this protein. In addition to platelets, TGF-beta1 is also found in other blood fractions, including plasma and the circulating leukocytes. However, more than 15 years after the initial isolation of TGF-beta1, there remains no consensus on how much TGF-beta1 is present in normal human plasma. Here we review the difficulties associated with measuring TGF-beta concentrations in complex biological fluids, and discuss the current state of knowledge on the distribution of TGF-beta isoforms in various blood fractions as well as the nature of the TGF-beta-containing protein complexes.

• Shah AH, Lee C
• TGF-beta-based immunotherapy for cancer: breaching the tumor firewall.
• Prostate. 2000; 45: 167-72
• Display abstract

• Many malignant cells secrete transforming growth factor-beta (TGF-beta), a potent immunosuppresant, suggesting that TGF-beta production may represent a significant tumor escape mechanism from host immunosurveillance. Establishment of a leukocyte subpopulation with disrupted TGF-beta signaling in the tumor-bearing host offers a potential means for immunotherapy of cancer. Downregulation of TGF-beta secretion in tumor cells results in restoration of immunogenicity in the host, while T-cell insensitivity to TGF-beta results in accelerated differentiation and autoimmunity, elements of which may be required in order to combat self-antigen-expressing tumors in a tolerized host. The rationale, approaches, and potential pitfalls of this strategy will be discussed.

• Choy L, Skillington J, Derynck R
• Roles of autocrine TGF-beta receptor and Smad signaling in adipocyte differentiation.
• J Cell Biol. 2000; 149: 667-82
• Display abstract

• TGF-beta inhibits adipocyte differentiation, yet is expressed by adipocytes. The function of TGF-beta in adipogenesis, and its mechanism of action, is unknown. To address the role of TGF-beta signaling in adipocyte differentiation, we characterized the expression of the TGF-beta receptors, and the Smads which transmit or inhibit TGF-beta signals, during adipogenesis in 3T3-F442A cells. We found that the cell-surface availability of TGF-beta receptors strongly decreased as adipogenesis proceeds. Whereas mRNA levels for Smads 2, 3, and 4 were unchanged during differentiation, mRNA levels for Smads 6 and 7, which are known to inhibit TGF-beta responses, decreased severely. Dominant negative interference with TGF-beta receptor signaling, by stably expressing a truncated type II TGF-beta receptor, enhanced differentiation and decreased growth. Stable overexpression of Smad2 or Smad3 inhibited differentiation and dominant negative inhibition of Smad3 function, but not Smad2 function, enhanced adipogenesis. Increased Smad6 and Smad7 levels blocked differentiation and enhanced TGF-beta-induced responses. The inhibitory effect of Smad7 on adipocyte differentiation and its cooperation with TGF-beta was associated with the C-domain of Smad7. Our results indicate that endogenous TGF-beta signaling regulates the rate of adipogenesis, and that Smad2 and Smad3 have distinct functions in this endogenous control of differentiation. Smad6 and Smad7 act as negative regulators of adipogenesis and, even though known to inhibit TGF-beta responses, enhance the effects of TGF-beta on these cells.

• Edens M, Leof EB
• In vitro assays for measuring TGF-beta growth stimulation and inhibition.
• Methods Mol Biol. 2000; 142: 1-11

• Muenke M, Cohen MM Jr
• Genetic approaches to understanding brain development: holoprosencephaly as a model.
• Ment Retard Dev Disabil Res Rev. 2000; 6: 15-21
• Display abstract

• Holoprosencephaly (HPE) is the most common major developmental defect of the forebrain in humans. Clinical expression is variable, ranging from a small brain with a single cerebral ventricle and cyclopia to clinically unaffected carriers in familial HPE. Significant etiologic heterogeneity exists in HPE and includes both genetic and environmental causes. Defects in the cell signaling pathway involving the Sonic Hedgehog (SHH) gene, as well as defects in the cholesterol biosynthesis have been shown to cause HPE in humans. More recently, HPE genes from additional signaling pathways have been identified. These discoveries and current genetic approaches serve as a paradigm for studying normal and abnormal brain morphogenesis. MRDD Research Reviews 6:15-21, 2000.

• Mazzieri R, Munger JS, Rifkin DB
• Measurement of active TGF-beta generated by cultured cells.
• Methods Mol Biol. 2000; 142: 13-27

• Danielpour D
• Iodination of TGF-beta, TGF-beta-receptor crosslinking, and immunoprecipitation of TGF-beta-receptor complexes.
• Methods Mol Biol. 2000; 142: 29-37

• King T, Brown NA
• Developmental biology. Antagonists on the left flank.
• Nature. 1999; 401: 222-3

• Boyer AS, Erickson CP, Runyan RB
• Epithelial-mesenchymal transformation in the embryonic heart is mediated through distinct pertussis toxin-sensitive and TGFbeta signal transduction mechanisms.
• Dev Dyn. 1999; 214: 81-91
• Display abstract

• During early development, progenitors of the heart valves and septa are formed by epithelial-mesenchymal transformation (EMT) of endothelial cells in the atrioventricular (AV) canal. Previously, we showed that pertussis toxin, a specific inhibitor of a subset of G proteins, inhibited EMT in chick AV canal cultures. This study examines in detail the effects of pertussis toxin on the process of EMT. One of the major mediators of EMT is Transforming Growth Factor beta 3 (TGFbeta3) which acts through the TGFbeta Type II receptor. To determine whether pertussis toxin affects EMT via the TGFbeta Type II receptor pathway, we compared AV cultures treated with pertussis toxin and TGFbeta Type II receptor blocking antibody. Pertussis toxin inhibited several elements of EMT. At all stages tested, pertussis toxin blocked endothelial cell-cell separation, cell hypertrophy, and the cellular polarization associated with endothelial activation. These activities were unaffected by TGFbeta Type II receptor antibodies. Pertussis toxin also reduced transformed mesenchymal cell migration by 61%. The expression patterns of several proteins (as markers of EMT) were analyzed in untreated, pertussis toxin-treated, and TGFbeta Type II receptor blocking antibody-treated cultures. These markers were alpha-smooth muscle actin, Mox-1, fibrillin 2, tenascin, cell surface beta 1,4 galactosyltransferase (GalTase), and integrin alpha6. Clear differences in marker expression were found between the two inhibitors. For example, in all cells, pertussis toxin inhibited expression of alpha-smooth muscle actin and GalTase while TGFbeta Type II receptor antibody treatment increased expression of these two proteins. These data suggest that G protein-mediated signaling is required for several elements of EMT. Furthermore, distinct G protein and TGFbeta signal transduction pathways mediate discrete components of EMT.

• Pujol JP
• TGF-beta and osteoarthritis: in vivo veritas?
• Osteoarthritis Cartilage. 1999; 7: 439-40

• Shibuya H
• [Functional role for TAB1-TAK1 in TGF-beta signaling]
• Seikagaku. 1999; 71: 1205-12

• Chen R, Halder G, Zhang Z, Mardon G
• Signaling by the TGF-beta homolog decapentaplegic functions reiteratively within the network of genes controlling retinal cell fate determination in Drosophila.
• Development. 1999; 126: 935-43
• Display abstract

• Retinal cell fate determination in Drosophila is controlled by an interactive network of genes, including eyeless, eyes absent, sine oculis and dachshund. We have investigated the role of the TGF-beta homolog decapentaplegic in this pathway. We demonstrate that, during eye development, while eyeless transcription does not depend on decapentaplegic activity, the expression of eyes absent, sine oculis and dachshund are greatly reduced in a decapentaplegic mutant background. We also show that decapentaplegic signaling acts synergistically with and at multiple levels of the retinal determination network to induce eyes absent, sine oculis and dachshund expression and ectopic eye formation. These results suggest a mechanism by which a general patterning signal such as Decapentaplegic cooperates reiteratively with tissue-specific factors to determine distinct cell fates during development.

• Bresalier RS
• Transforming growth factor beta and tumor-suppressing pathways: it doesn't matter how you get there, as long as you get there.
• Gastroenterology. 1999; 117: 500-2

• Lau A, Matzuk MM
• Transgenic models in the study of reproduction.
• Gravit Space Biol Bull. 1999; 12: 75-82
• Display abstract

• The development of techniques to manipulate genes within mouse embryonic stem (ES) cells has allowed investigators to study the functions of many genes in vivo. We have used these techniques to functionally mutate genes to study how the loss of the gene affects development, oncogenesis, and reproduction. Genes affecting development include members of the transforming growth factor-beta (TGF-beta) superfamily and their signaling pathway. We have shown that mutations in this complicated signaling network affect a wide range of embryonic developmental processes including craniofacial morphogenesis, dentition and muscle development. One specific member of TGF-beta family, inhibin alpha, has been identified as a novel tumor suppressor in the testes, ovaries, and adrenal glands. Another focus of research in the laboratory is the analysis of roles of proteins in the hypothalamic-pituitary-gonadal axis and the affect of disrupting this pathway on reproductive function. We have demonstrated that several genes expressed in the pituitary and gonads are required for folliculogenesis leading to female infertility and in two cases are important for Sertoli cell proliferation in males. The studies using ES cell technology has enabled us to dissect two complex networks in animal models.

• Kim YS, Yi Y, Choi SG, Kim SJ
• Development of TGF-beta resistance during malignant progression.
• Arch Pharm Res. 1999; 22: 1-8
• Display abstract

• Transforming growth factor-beta (TGF-beta) is the prototypical multifunctional cytokine, participating in the regulation of vital cellular activities such as proliferation and differentiation as well as a number of basic physiological functions. The effects of TGF-beta are critically dependent on the expression and distribution of a family of TGF-beta receptors, the TGF-beta types I, II, and III. It is now known that a wide variety of human pathology can be caused by aberrant expression and function of these receptors. The coding sequence of the type II receptor (RII) appears to render it uniquely susceptible to DNA replication errors in the course of normal cell division. By virtue of its key role in the regulation of cell proliferation, TGF-beta RII should be considered as a tumor suppressor gene. High levels of mutation in the TGF-beta RII gene have been observed in a wide range of primarily epithelial malignancies, including colon and gastric cancer. It appears likely that mutation of the TGF-beta RII gene may be a very critical step in the pathway of carcinogenesis.

• Ring CJ, Cho KW
• Specificity in transforming growth factor-beta signaling pathways.
• Am J Hum Genet. 1999; 64: 691-7

• Kelly DL, Rizzino A
• Growth regulatory factors and carcinogenesis: the roles played by transforming growth factor beta, its receptors and signaling pathways.
• Anticancer Res. 1999; 19: 4791-807
• Display abstract

• It is widely recognized that growth factors play critical roles in cell proliferation and differentiation. In the early 1980s, several members of the transforming growth factor-beta (TGF-beta) superfamily were identified and subsequently shown to play important roles in many diseases, in particular cancer. Efforts to understand how TGF-beta exerts its effects led to identification of TGF-beta-receptors and several downstream signaling pathways activated by this family of growth factors. This review provides an overview of TGF-beta-receptors and its downstream signaling pathways. As part of this discussion, this review indicates that inactivation of TGF-beta-receptors or components of their signaling pathways is often a target during carcinogenesis and that mutations or altered expression at any step along this complex, growth regulatory pathway can lead to aberrant cell proliferation. Lastly, the Cancer Genome Anatomy Project is briefly discussed, in particular how it may help to identify aberrant growth factor expression during carcinogenesis and improve the diagnosis of cancer patients.

• Sporn MB
• TGF-beta: 20 years and counting.
• Microbes Infect. 1999; 1: 1251-3
• Display abstract

• The history of transforming growth factor-beta (TGF-beta) as a bifunctional agent in the immune system is briefly described. The importance of cellular context in understanding the role of TGF-beta in regulating immune response is emphasized.

• Khalil N
• TGF-beta: from latent to active.
• Microbes Infect. 1999; 1: 1255-63
• Display abstract

• The transforming growth factor-betas (TGF-betas) are synthesized as precursor proteins that are modified intracellularly prior to secretion. One of the most relevant intracellular modifications is the cleavage of the C-terminal pro-region from the N-terminal portion of the protein. The C-terminal pro-region is referred to as the latency-associated peptide (LAP) while the N-terminal region is called the mature TGF-beta or active TGF-beta. However, with some exceptions the LAP noncovalently associates with the mature TGF-beta prior to secretion. When the mature TGF-beta is associated with the LAP it is called L-TGF-beta and cannot interact with its receptor and has no biological effect. The TGF-betas and their receptors are very ubiquitously expressed, suggesting that the regulation of TGF-beta activity is likely to be complex and multifactorial. However, one of the most important means of controlling the biological effects of TGF-beta is the regulation of converting L-TGF-beta to active TGF-beta. The current literature supports two major mechanisms of activation of L-TGF-beta and suggests that the mechanism of activation of L-TGF-beta may be varied and context-dependent. For TGF-beta to become biologically active the LAP has to be either released from its associations with L-TGF-beta or undergo conformational change such that the LAP is not released from the L-TGF-beta complex but exposes the TGF-beta receptor binding site. Since TGF-beta has been associated with the pathogenesis of numerous diseases, the various mechanisms of activation of L-TGF-beta in context offer the possibility of controlling TGF-beta activity localized to the organ of involvement and to a more specific disease process.

• Bonewald LF
• Regulation and regulatory activities of transforming growth factor beta.
• Crit Rev Eukaryot Gene Expr. 1999; 9: 33-44
• Display abstract

• Transforming growth factor beta (TGF beta) is unique among growth factors in its potent and widespread actions. Almost every cell in the body has been shown to make some form of TGF beta, and almost every cell has receptors for TGF beta. Therefore, it becomes apparent that this growth factor must be tightly regulated to prevent disease. The mechanisms of regulation of TGF beta are extensive and complex. One set of mechanisms centers around the fact that TGF beta is produced in a latent form that must be activated to produce biologically active TGF beta. These mechanisms include the latency of the molecule, the production of various latent forms, its targeting to cells for activation or to matrix for storage, and the means of activation of the latent forms. The TGF beta isoforms and the types, affinity, and signaling functions of its receptors also add complexity to the regulation of the effects of TGF beta. Active TGF beta regulates numerous processes in the body. TGF beta has three major biological effects: growth inhibition, stimulation of extracellular matrix formation, and immunosuppression. The means by which TGF beta regulates the expression of the numerous genes on which it has effects are complex and more information is needed. The means by which TGF beta regulates gene expression and the means by which the actions of TGF beta are regulated are addressed in this review.

• Gold LI, Parekh TV
• Loss of growth regulation by transforming growth factor-beta (TGF-beta) in human cancers: studies on endometrial carcinoma.
• Semin Reprod Endocrinol. 1999; 17: 73-92
• Display abstract

• Members of the Transforming Growth Factor-beta (TGF-beta) family are one of the few endogenous inhibitors of cell growth. As uncontrolled cellular proliferation is a hallmark of cancer, an important question to address is how cancer cells escape normal growth regulatory mechanisms to become malignant. In this context, components of the TGF-beta growth response pathway are considered to be tumor suppressor genes, as absence of one or more of TGF-beta receptor and signaling proteins cause loss of cell growth regulation through an inability to regulate proteins that directly block cells in G1 phase of the cell cycle. Endometrial carcinoma (ECA) provides an excellent paradigm to study the changes that accompany loss of TGF-beta-mediated growth, control as a function of neoplastic development, since it is generally preceded by complex hyperplasia. Type 1 ECA is characterized as an estrogen-induced cancer, which responds well to progestin therapy. Since it has become increasingly evident that steroids can regulate growth through growth factors, ECA is also an ideal model for investigating the role for gonadal steroids in the loss of TGF-beta growth regulation in the etiopathogenesis of ECA. Thus, hormonal carcinogenesis adds another level of complexity in studying loss of growth regulation in human cancers. The purpose of this review is to 1) provide the most current background information on how TGF-beta functions including its activation, receptors, signal transduction mechanisms, and control of the cell cycle. 2) present recent information that shows how malignant cells subvert the growth inhibitory effects of TGF-beta by incurring defects in every aspect of the pathway that mediates the TGF-beta growth inhibitory response, and 3) describe the putative role for TGF-beta in the oncogenesis of ECA, provided primarily by the results from our laboratory. Understanding the molecular events involved in TGF-beta function in normal cells and its lack of function in tumor cells should identify novel therapeutic targets in human cancers.

• Roberts AB
• TGF-beta signaling from receptors to the nucleus.
• Microbes Infect. 1999; 1: 1265-73
• Display abstract

• In the past three years, a novel signal transduction pathway downstream of the transforming growth factor-beta (TGF-beta) superfamily receptor serine-threonine kinases has been shown to be mediated by a family of latent transcription factors called 'Smads'. These proteins mediate a short-circuited pathway in which a set of receptor-activated Smads are phosphorylated directly by the receptor kinase and then translocate to the nucleus complexed to the common mediator, Smad4, to participate in transcriptional complexes. Smads 2 and 3 mediate signals predominantly from the TGF-beta receptors. Of these, specific roles have been ascribed to Smad3 in control of chemotaxis of neutrophils and macrophages and the inhibition of Smad3 activity by the oncogene Evi-1 suggests that it may play a role in leukemogenesis. Other data, such as the induction by the inflammatory cytokine interferon-gamma of an inhibitory Smad, Smad7, which blocks the actions of Smad3, suggest that identification of the specific gene targets of Smad proteins in immune cells will provide new insight into the mechanisms of TGF-beta action on these cells.

• Miyazono K
• [Receptors and signal transduction for the TGF-beta related factors]
• Nippon Ronen Igakkai Zasshi. 1999; 36: 162-6

• Cerwenka A, Swain SL
• TGF-beta1: immunosuppressant and viability factor for T lymphocytes.
• Microbes Infect. 1999; 1: 1291-6
• Display abstract

• Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine with multiple roles in the immune system. To date, it has been difficult to develop a comprehensive picture of the effect of TGF-beta on T lymphocytes, because TGF-beta not only acts directly on T lymphocytes, but also acts indirectly by regulating the function of antigen-presenting cells. In early studies, it was mostly the inhibitory function of TGF-beta that was demonstrated; recently, however TGF-beta was recognized as an antiapoptotic survival factor for T lymphocytes. The outcome of the TGF-beta effect on T lymphocytes was shown to strongly depend on their stage of differentiation and on the cytokine milieu. TGF-beta cannot be classified as a classical Th1 or Th2 cytokine. However, recently the existence of the TGF-beta-producing Th3 subset was described which might play an important regulatory role during an immune response. A better understanding of the molecular mechanism of how TGF-beta inhibits or stimulates T lymphocytes will help to predict the complex functions of this cytokine.

• Kawabata M, Miyazono K
• Signal transduction of the TGF-beta superfamily by Smad proteins.
• J Biochem (Tokyo). 1999; 125: 9-16
• Display abstract

• Members of the TGF-beta superfamily regulate the growth and differentiation of various types of cells. Smads are recently identified proteins that mediate intracellular signaling of the TGF-beta superfamily. Smads are grouped into three classes depending on their structure and functions. R-Smads are phosphorylated by type I serine-threonine kinase receptors for TGF-beta superfamily members. R-Smads then associate with Co-Smads. Smad4 is the only vertebrate Co-Smad identified thus far, and is required for the signaling pathways of different ligands. The heteromeric Smad complex translocates into the nucleus, where it activates target genes. Anti-Smads inhibit signaling by R-Smads and Co-Smads. Smads bind to DNA directly or indirectly via other DNA binding proteins. R-Smads interact with transcriptional coactivators, and have intrinsic transactivation activity. Elucidation of the functions of Smads will provide the framework for research on TGF-beta superfamily signaling.

• Raftery LA, Sutherland DJ
• TGF-beta family signal transduction in Drosophila development: from Mad to Smads.
• Dev Biol. 1999; 210: 251-68
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily encompasses a large group of soluble extracellular proteins that are potent regulators of development in both vertebrates and invertebrates. Drosophila TGF-beta family members include three proteins with homology to vertebrate bone morphogenetic proteins (BMPs): Decapentaplegic (Dpp), Screw, and Glass bottom boat-60A. Genetic studies of Dpp signaling led to the identification of Smad proteins as central mediators of signal transduction by TGF-beta family members. Work in mammalian tissue culture has elucidated a biochemical model for signal transduction, in which activation of receptor serine-threonine kinase activity leads to phosphorylation of specific Smad proteins and translocation of heteromeric Smad protein complexes to the nucleus. Once in the nucleus Smad proteins interact with other DNA binding proteins to regulate transcription of specific target genes. Dissection of Dpp-response elements from genes expressed during embryonic mesoderm patterning and midgut morphogenesis provides important insights into the contributions of Smad proteins and tissue-specific transcription factors to spatial regulation of gene expression. Genetic studies in Drosophila are now expanding to include multiple BMP ligands and receptors and have uncovered activities not explained by the current signal transduction model. Identification of more ligand sequences and demonstration of a functional Drosophila activin-like signal transduction pathway suggest that all TGF-beta signal transduction pathways are present in flies.

• Vogel G
• A new blocker for the TGF-beta pathway.
• Science. 1999; 286: 665-665

• Mummery CL, van den Eijnden-van Raaij AJ
• Developmental tumours, early differentiation and the transforming growth factor beta superfamily.
• Int J Dev Biol. 1999; 43: 693-709
• Display abstract

• Embryonal carcinoma and embryonic stem cells have been very useful models for identifying some of the factors that regulate differentiation in early mammalian development. Here, we present a brief history of their original isolation and characterization and of their later introduction into the Hubrecht Laboratory. We illustrate in a review their contribution to our current understanding of the function of transforming growth factor beta and ligands binding to the receptors of a related factor, activin, in development with some of our own work.

• Padgett RW
• Intracellular signaling: Fleshing out the TGFbeta pathway.
• Curr Biol. 1999; 9: 40811-40811
• Display abstract

• Recent studies of the intracellular signaling pathway initiated by ligands of the transforming growth factor beta (TGFbeta) family have provided new insights into how the receptors for such ligands phosphorylate their substrates - the Smads - and how signaling specificity is achieved.

• Reiss M
• TGF-beta and cancer.
• Microbes Infect. 1999; 1: 1327-47
• Display abstract

• The relationships between transforming growth factor-beta (TGF-beta) and cancer are varied and complex. The paradigm that is emerging from the experimental evidence accumulated over the past decade or so is that TGF-beta can play two different and opposite roles with respect to the process of malignant progression. During early stages of carcinogenesis, TGF-beta acts predominantly as a potent tumor suppressor and may mediate the actions of chemopreventive agents such as retinoids and nonsteroidal anti-estrogens. However, at some point during the development and progression of malignant neoplasms, bioactive TGF-betas make their appearance in the tumor microenvironment and the tumor cells escape from TGF-beta-dependent growth arrest. In many cases, this resistance to TGF-beta is the consequence of loss or mutational inactivation of the genes that encode signaling intermediates. These include the types I and II TGF-beta receptors, as well as receptor-associated and common-mediator Smads. The stage of tumor development or progression at which TGF-beta-resistant clones come to dominate the tumor cell population in different types of neoplasm remains to be defined. The phenotypic switch from TGF-beta-sensitivity to TGF-beta-resistance that occurs during carcinogenesis has several important implications for cancer prevention and treatment.

• Wahl SM
• TGF-beta in the evolution and resolution of inflammatory and immune processes. Introduction.
• Microbes Infect. 1999; 1: 1247-9

• ten Dijke P, Heldin CH
• Signal transduction. An anchor for activation.
• Nature. 1999; 397: 109111-109111

• Duff EK, Clarke AR
• Smad4 (DPC4)--a potent tumour suppressor?
• Br J Cancer. 1998; 78: 1615-9
• Display abstract

• The recently described family of Smad molecules are essential mediators of transforming growth factor beta (TGF-beta) signalling. To date, seven members of this family have been identified, each of which plays a specific and separate role in mediating TGF-beta superfamily gene transcription. At least two different Smads, Smad2 and Smad4 (DPC4), have been implicated in human cancer and appear to have tumour-suppressor functions. Loss of function of Smad4 is most strongly associated with human pancreatic and colorectal malignancy. Furthermore, work from several different groups has suggested associations between Smad4 loss and malignancy in a number of other tissues. Here, we present a review of the current state of the literature implicating the central Smad mediator, Smad4, in the development of cancer.

• Persson U et al.
• The L45 loop in type I receptors for TGF-beta family members is a critical determinant in specifying Smad isoform activation.
• FEBS Lett. 1998; 434: 83-7
• Display abstract

• Transforming growth factor-beta (TGF-beta) and bone morphogenetic proteins (BMPs) signal via distinct type I and type II receptors and Smad proteins. A nine amino acid sequence between kinase subdomains IV and V in type I receptors, termed the L45 loop, has been shown to be important in conferring signalling specificity. We examined the responses of a mutant TGF-beta type I receptor (TbetaR-I) and a mutant BMPR-IB, in which the L45 regions of these two receptors were exchanged. Swapping the four amino acid residues that are different in BMPR-IB for those in TbetaR-I, and vice versa, switched their type I receptor-restricted Smad activation and specificity in transcriptional responses. These studies identify the L45 loop regions in type I receptors as critical determinants in specifying Smad isoform activation.

• Letterio JJ, Bottinger EP
• TGF-beta knockout and dominant-negative receptor transgenic mice.
• Miner Electrolyte Metab. 1998; 24: 161-7
• Display abstract

• Use of homologous recombination and transgenic technologies have provided mouse models to study the physiological roles of the three mammalian TGF-beta isoforms, and their regulation in the context of the intact animal. Mice harboring null mutations for TGF-beta isoforms demonstrate that each exerts discrete nonoverlapping functions during development. TGF-beta1 null mice reveal a crucial role for this cytokine in modulation of the immune system, with evidence for altered development, activation and function of various immune cell populations. New approaches to tissue- and cell-restricted disruption of TGF-beta signaling pathways in transgenic mice carrying dominant-negative mutant TGF-beta receptors will be discussed.

• Padgett RW, Cho SH, Evangelista C
• Smads are the central component in transforming growth factor-beta signaling.
• Pharmacol Ther. 1998; 78: 47-52
• Display abstract

• Until recently, little was known about how transforming growth factor (TGF)-beta signals are transduced to the nucleus. With the discovery of the Smad proteins initially in Drosophila and C. elegans, the unraveling of the pathway has begun. Nine different vertebrate members also have been reported, indicating that Smads are a conserved component of the TGF-beta pathway. Currently, there are three functional classes of Smads. Class I Smads are phosphorylated by TGF-beta receptors and move to the nucleus. The Class II Smads function with Class I Smads, while Class III Smads antagonize the function of Class I Smads. New evidence shows that Smads bind specific DNA sequences and induce transcription of downstream target genes, thus placing the Smads at the center of the TGF-beta signaling pathway.

• Clark DA, Coker R
• Transforming growth factor-beta (TGF-beta).
• Int J Biochem Cell Biol. 1998; 30: 293-8
• Display abstract

• The TGF-beta family of cytokines are ubiquitous, multifunctional and essential to survival. They play important roles in growth and development, inflammation and repair and host immunity. The mammalian TGF-beta isoforms (TGF-beta 1, beta 2 and beta 3) are secreted as latent precursors and have multiple cell surface receptors of which at least two mediate signal transduction. Autocrine and paracrine effects of TGF-beta's can be modified by extracellular matrix, neighbouring cells and other cytokines. The vital role of the TGF-beta family is illustrated by the fact that approximately 50% of TGF-beta 1 gene knockout mice die in utero and the remainder succumb to uncontrolled inflammation after birth. TGF-beta 2 and TGF-beta 3 gene knockout mice are not yet described. More recently, novel TGF-beta-like molecules have been described which share some of the properties of the mammalian TGF-beta isoforms. The role of TGF-beta in homeostatic and pathogenic processes suggests numerous applications in the diagnosis and treatment of various diseases characterised by inflammation and fibrosis.

• Hu PP, Datto MB, Wang XF
• Molecular mechanisms of transforming growth factor-beta signaling.
• Endocr Rev. 1998; 19: 349-63

• Padgett RW, Das P, Krishna S
• TGF-beta signaling, Smads, and tumor suppressors.
• Bioessays. 1998; 20: 382-90
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily is used throughout animal development for regulating the growth and patterning of many tissue types. During the past few years, rapid progress has been made in deciphering how TGF-beta signals are transduced from outside the cell to the nucleus. This progress is based on biochemical studies in vertebrate systems and a combination of genetic studies in Drosophila and Caenorhabditis elegans. These studies have identified a novel family of signaling proteins, the Smad family. Smads can act positively and be phosphorylated by TGF-beta-like receptors or can act negatively and prevent activation of the positively acting group. The positively acting Smads translocate to the nucleus, bind DNA, and act as transcriptional activators. Thus, genetic and biochemical studies suggest a very simple signaling pathway, in which Smads are the primary downstream participant.

• Attisano L, Wrana JL
• Mads and Smads in TGF beta signalling.
• Curr Opin Cell Biol. 1998; 10: 188-94
• Display abstract

• The discovery of the Mothers against dpp (Mad) gene in Drosophila has opened a window on an entirely unique signalling pathway that functions to mediate responses to the tumour growth factor beta (TGF beta) superfamily. This pathway, which is comprised of a family of proteins related to Mad, acts to convey signals directly from TGF beta receptors to the nucleus and is implicated in the pathogenesis of human diseases.

• Nishihara A et al.
• Role of p300, a transcriptional coactivator, in signalling of TGF-beta.
• Genes Cells. 1998; 3: 613-23
• Display abstract

• BACKGROUND: Smad proteins are novel transcriptional regulators mediating the signalling of the transforming growth factor-beta (TGF-beta) superfamily. Coactivators such as p300/CBP promote transactivation by various transcription factors through a direct interaction with them. Adenoviral oncoprotein E1A, which binds p300, was shown to inhibit the signalling of TGF-beta. These findings raise the possibility that p300 may be involved in TGF-beta signalling. RESULTS: We investigated whether p300 is involved in transactivation by Smads. p300 enhanced the Smad-induced transactivation of p3TP-Lux, a TGF-beta responsive reporter. E1A inhibited this enhancement, and the inhibition required its ability to bind p300/CBP. p300 and Smad3, as well as Smad2, interacted in vivo in a ligand-dependent manner. The binding region in Smad3 was its C-terminal half that was previously shown to possess an intrinsic transactivation activity. The binding region in p300 was mapped to its C-terminal 678 amino acids. The minimal Smad2/3-interacting region, as well as the rest of the p300, inhibited the transactivation of p3TP-Lux in a dominant-negative fashion. CONCLUSION: p300 interacted with Smad2 and Smad3 in a ligand-dependent manner, and enhanced the transactivation by Smads. Our results present the molecular basis of the transactivation by Smad proteins.

• Blader P, Strahle U
• Casting an eye over cyclopia.
• Nature. 1998; 395: 112-3

• Ahmed W et al.
• Signaling pathway by which TGF-beta1 increases expression of latent TGF-beta binding protein-2 at the transcriptional level.
• Connect Tissue Res. 1998; 37: 263-76
• Display abstract

• The cytokine transforming growth factor-beta has multiple effects on a wide variety of cell types. These effects include modulation of growth and regulation of gene transcription. In the present work, we demonstrate that TGF-beta1 increases transcription of the latent transforming growth factor-beta binding protein-2 ( LTBP-2) gene in cultured human fetal lung fibroblasts leading to a significant increase in LTBP-2 mRNA steady state level. The stability of LTBP-2 mRNA was not appreciably altered. A corresponding increase in production of LTBP-2 protein accompanied the increase in mRNA. Through the use of specific inhibitors, we demonstrate that a member of the Ras super family and a protein kinase C, probably of the atypical (non-diacylglycerol, non-Ca++ dependent) class are likely to be components in the signaling pathway. However, phospholipases, G proteins and extracellular-signal regulated kinases do not appear to be involved. These results combined with previous findings on elastin regulation by TGF-beta1 (Kucich et al. (1997). Am. J. Respir. Cell Mol. Biol., 17: 10-16) demonstrate that TGF-beta1 can coordinately increase the steady state levels of mRNAs encoding components of the elastic fiber, but through diverse mechanisms. In contrast to LTBP-2, increased elastin expression is achieved by message stabilization. Furthermore, the TGF-beta1 signaling pathways differ and while the pathway leading to increased LTBP-2 transcription shares components with those modulating transcription of other genes, it is unlikely to be precisely congruent with any other previously described one.

• Cho KW, Blitz IL
• BMPs, Smads and metalloproteases: extracellular and intracellular modes of negative regulation.
• Curr Opin Genet Dev. 1998; 8: 443-9
• Display abstract

• Bone morphogenetic (BMPs), members of the TGF-beta superfamily, have critical functions in many biological contexts. Recent findings in Drosophila and vertebrates suggest that BMP signaling can be modulated extracellularly and intracellularly by the availability of BMP inhibitors and Smads, respectively.

• Massague J
• TGF-beta signal transduction.
• Annu Rev Biochem. 1998; 67: 753-91
• Display abstract

• The transforming growth factor beta (TGF-beta) family of growth factors control the development and homeostasis of most tissues in metazoan organisms. Work over the past few years has led to the elucidation of a TGF-beta signal transduction network. This network involves receptor serine/threonine kinases at the cell surface and their substrates, the SMAD proteins, which move into the nucleus, where they activate target gene transcription in association with DNA-binding partners. Distinct repertoires of receptors, SMAD proteins, and DNA-binding partners seemingly underlie, in a cell-specific manner, the multifunctional nature of TGF-beta and related factors. Mutations in these pathways are the cause of various forms of human cancer and developmental disorders.

• Dobens LL, Raftery LA
• Drosophila oogenesis: a model system to understand TGF-beta/Dpp directed cell morphogenesis.
• Ann N Y Acad Sci. 1998; 857: 245-7

• Visser JA, Themmen AP
• Downstream factors in transforming growth factor-beta family signaling.
• Mol Cell Endocrinol. 1998; 146: 7-17
• Display abstract

• The recently identified family of Smad proteins has given insight in the understanding of how members of the transforming growth factor-beta (TGF-beta) family relay their signal to the nucleus. Besides Smad proteins, G proteins and MAPKs are also involved in the downstream signaling of TGF-beta family members. The identification of elements that function downstream in the TGF-beta signaling pathway and the fact that these downstream players can interact with the signaling cascade of other growth factors, may give insight into the diverse biological responses evoked by the TGF-beta family members.

• Chen YG et al.
• Determinants of specificity in TGF-beta signal transduction.
• Genes Dev. 1998; 12: 2144-52
• Display abstract

• Signal transduction by the TGF-beta family involves sets of receptor serine/threonine kinases, Smad proteins that act as receptor substrates, and Smad-associated transcription factors that target specific genes. We have identified discrete structural elements that dictate the selective interactions between receptors and Smads and between Smads and transcription factors in the TGF-beta and BMP pathways. A cluster of four residues in the L45 loop of the type I receptor kinase domain, and a matching set of two residues in the L3 loop of the Smad carboxy-terminal domain establish the specificity of receptor-Smad interactions. A cluster of residues in the highly exposed alpha-helix 2 of the Smad carboxy-terminal domain specify the interaction with the DNA-binding factor Fast1 and, as a result, the gene responses mediated by the pathway. By establishing specific interactions, these determinants keep the TGF-beta and BMP pathways segregated from each other.

• Toossi Z, Ellner JJ
• The role of TGF beta in the pathogenesis of human tuberculosis.
• Clin Immunol Immunopathol. 1998; 87: 107-14

• Hata A, Shi Y, Massague J
• TGF-beta signaling and cancer: structural and functional consequences of mutations in Smads.
• Mol Med Today. 1998; 4: 257-62
• Display abstract

• Transforming growth factor-beta (TGF-beta) and related cytokines control the development and homeostasis of many tissues by regulating the expression of genes that determine cell phenotype. Recent progress has elucidated the way in which members of the TGF-beta family initiate their signal through transmembrane receptors and transmit it to target genes via the Smad family of signal-transducing proteins. This review describes TGF-beta signaling pathways as currently understood and mutations of the genes that encode Smads that disrupt the function of these proteins and cause various forms of cancer.

• Kurokawa M et al.
• The oncoprotein Evi-1 represses TGF-beta signalling by inhibiting Smad3.
• Nature. 1998; 394: 92-6
• Display abstract

• Evi-1 encodes a zinc-finger protein that may be involved in leukaemic transformation of haematopoietic cells. Evi-1 has two zinc-finger domains, one with seven repeats of a zinc-finger motif and one with three repeats, and it has characteristics of a transcriptional regulator. Although Evi-1 is thought to be able to promote growth and to block differentiation in some cell types, its biological functions are poorly understood. Here we study the mechanisms that underlie oncogenesis induced by Evi-1 by investigating whether Evi-1 perturbs signalling through transforming growth factor-beta (TGF-beta), one of the most studied growth-regulatory factors, which inhibits proliferation of a wide range of cell types. We show that Evi-1 represses TGF-beta signalling and antagonizes the growth-inhibitory effects of TGF-beta. Two separate regions of Evi-1 are responsible for this repression; one of these regions is the first zinc-finger domain. Through this domain, Evi-1 interacts with Smad3, an intracellular mediator of TGF-beta signalling, thereby suppressing the transcriptional activity of Smad3. These results define a new function of Evi-1 as a repressor of signalling through TGF-beta.

• Bottinger EP, Miao J, Schiffer M, Bitzer M, Roberts IS
• Transforming growth factor beta signal transduction in the kidney.
• Kidney Blood Press Res. 1998; 21: 259-61

• Neul JL, Ferguson EL
• Spatially restricted activation of the SAX receptor by SCW modulates DPP/TKV signaling in Drosophila dorsal-ventral patterning.
• Cell. 1998; 95: 483-94
• Display abstract

• Dorsal-ventral patterning within the embryonic ectoderm of Drosophila requires two TGFbeta ligands, DPP and SCW, and two type I TGFbeta receptors, TKV and SAX. In embryos lacking dpp signaling, increasing the level of TKV activity promotes progressively more dorsal cell types, while activation of SAX alone has no phenotypic consequences. However, SAX activity synergizes with TKV activity to promote dorsal development. Functional experiments suggest the two receptors have different ligands: DPP acts through TKV, and SCW acts through SAX. Furthermore, SOG, a negative regulator of this patterning process, preferentially blocks SCW activity. We propose that spatial regulation of the SAX pathway modulates TKV signaling to create positional information over the embryonic ectoderm.

• McCartney-Francis NL, Frazier-Jessen M, Wahl SM
• TGF-beta: a balancing act.
• Int Rev Immunol. 1998; 16: 553-80
• Display abstract

• Regulation of developmental processes as well as host defense and repair mechanisms requires the maintenance of a delicate balance of positive and negative regulatory signals. TGF-beta, a molecule known for its many diverse activities, can promote or inhibit cell growth and function. Disruption of the balance between these opposing activities can contribute to aberrant development, malignancy, or pathogenic immune and inflammatory responses. TGF-beta transgenic mouse studies highlight the essential function(s) of TGF-beta and its receptors and provide insight to potential therapeutic approaches to manipulate TGF-beta expression.

• Bitzer M, Sterzel RB, Bottinger EP
• Transforming growth factor-beta in renal disease.
• Kidney Blood Press Res. 1998; 21: 1-12
• Display abstract

• An extensive number of animal and clinical studies indicate that transforming growth factor-beta (TGF-beta s) play an important role in inflammatory and fibrotic diseases, including renal fibrosis. Recent mouse models harboring genetically engineered alterations in TGF-beta pathways reveal complicated mechanisms of regulation of TGF-beta activity in vivo. The purpose of this review is to present recent advances relevant to our understanding of the TGF-beta-signaling system in renal physiology and pathophysiology.

• Wrana JL
• TGF-beta receptors and signalling mechanisms.
• Miner Electrolyte Metab. 1998; 24: 120-30
• Display abstract

• Transforming growth factor-beta (TGF-beta) is the founding member of a large superfamily of related growth and differentiation factors that include bone morphogenetic proteins and activins. TGF-beta signals through two related transmembrane ser/thr kinase receptors, the type I and type II receptors. Signalling is initiated when the ligand binds to the type II receptor which is followed by recruitment of the type I receptor into a heteromeric complex. Within the complex the type II receptor transphosphorylates and activates the type I receptor kinase which targets downstream signalling components of the pathway. Proteins related to the Drosophila gene Mothers against dpp (MAD) are critical downstream substrates of the type I kinase. The vertebrate members of the MAD-related family, termed Smad2 and Smad3, interact specifically with the TGF-beta type I receptor and are phosphorylated on the last two serines of a conserved C-terminal SSXS motif. This phosphorylation induces association between these receptor-regulated Smads and Smad4 followed by translocation of the heteromeric complex to the nucleus. In the nucleus, heteromeric complexes of Smads can interact with DNA and with specific DNA binding transcription factors to elicit gene responses to TGF-beta. Thus TGF-beta signalling involves a direct pathway from the cell surface receptors to the nucleus. Recently, a novel mechanism to negatively regulate TGF-beta signalling was described that involves another class of MADR proteins. These anti-MADR proteins potently inhibit TGF-beta signalling by functioning as direct antagonists of the TGF-beta receptor type I kinase domain.

• Perrella MA, Jain MK, Lee ME
• Role of TGF-beta in vascular development and vascular reactivity.
• Miner Electrolyte Metab. 1998; 24: 136-43
• Display abstract

• Transforming growth factor-beta (TGF-beta) is the prototypic member of a large family of structurally related proteins. Three vertebrate TGF-beta isoforms have been identified and termed TGF-beta1, TGF-beta2, and TGF-beta3, respectively. In addition, two receptors of the serine/threonine kinase family termed type I and II have also been identified. In this review, we focused our attention on the effects of TGF-beta on vascular development and vascular reactivity. The critical role of the TGF-beta1 and the TGF-beta type II receptor in blood vessel formation in the yolk sac has been demonstrated by gene deletion experiments. Recent investigations have also shown that isoforms of TGF-beta play a critical role in smooth muscle cell differentiation. And, finally, a role for TGF-beta1 in the regulation of vascular tone and reactivity has been suggested by studies demonstrating that TGF-beta1 can inhibit the production of potent vasodilators (such as nitric oxide) and stimulate the production of potent vasoconstrictors (such as endothelin). Taken together, these studies suggest that TGF-beta plays a critical role in blood vessel development and vascular function.

• Ando T, Okuda S, Yanagida T, Fujishima M
• Localization of TGF-beta and its receptors in the kidney.
• Miner Electrolyte Metab. 1998; 24: 149-53
• Display abstract

• This paper reviews the expression and localization of proteins of the TGF-beta system in the kidney, i.e. TGF-beta isoforms, latent TGF-beta binding protein, and receptors. There is heterogeneity of TGF-beta isoforms, binding proteins and receptors. TGF-betas are more evenly distributed among different segments of the nephron than TGF-beta receptors, and are expressed by almost all cell types in the kidney. The spatial distribution of TGF-beta and its signaling receptor may help to better understand the biologic function of TGF-beta in the kidney.

• Bottinger EP, Kopp JB
• Lessons from TGF-beta transgenic mice.
• Miner Electrolyte Metab. 1998; 24: 154-60
• Display abstract

• Aberrant expression of TGF-beta and/or receptor/signaling function is present in a wide variety of disease processes. Overexpression of TGF-beta isoforms in transgenic mice using tissue-specific promoters has provided model systems to study the effects of increased activity of TGF-beta in the intact organism. We will review the pertinent features of some of these models, and discuss new insights provided by these studies into regulation and role of TGF-beta in health and disease.

• Heberden C, Denis I, Pointillart A, Mercier T
• TGF-beta and calcitriol.
• Gen Pharmacol. 1998; 30: 145-51
• Display abstract

• Vitamin D3 and transforming growth factor-beta (TGF-beta) are molecules from unrelated families that share identical actions on cell growth and differentiation. The active metabolite of vitamin D3, calcitriol (1alpha,25-dihydroxyvitamin D3), induces an inhibitory effect on the growth of various cell types, and the expression of different markers of cell differentiation. As the receptor of vitamin D3 is ubiquitous, these effects are widespread in the organism. TGF-beta is a growth factor produced by many cell types, and is a known inhibitor of the proliferation of epithelial cells. Because of the similarity in their actions, many studies have been aimed at defining some interactions between the two substances. The purpose of this article is to illustrate the nature of the interactions, and two examples are developed. In normal or transformed epithelial cells, it has been demonstrated that the inhibitory effect of calcitriol on cell growth could be related to an induction of TGF-beta synthesis, and of a paracrine/autocrine loop. In bone, where both compounds play a very important role on the mechanisms controlling bone formation and remodeling, the interplay is more complex, and even includes the receptors of the two substances. Interest in this topic is growing and will surely lead to the establishment of new links between those two compounds.

• Yanagisawa K et al.
• Induction of apoptosis by Smad3 and down-regulation of Smad3 expression in response to TGF-beta in human normal lung epithelial cells.
• Oncogene. 1998; 17: 1743-7
• Display abstract

• Smad family members are essential intracellular signaling components of the transforming growth factor-beta (TGF-beta) superfamily involved in a range of biological activities. Two highly homologous molecules, Smad2 and Smad3, have so far been identified as receptor-activated Smads for TGF-beta signaling and have become the focus of intensive studies. However, no definite differences in regulation or function have been established between these TGF-beta signaling molecules. In the present study, we show that the expression of Smad3, but not its close relative, Smad2, is down-regulated by TGF-beta mediated signals themselves in human lung epithelial cells. This down-regulation of Smad3 by TGF-beta treatment did not appear to result from shortening of the half-life of Smad3 mRNA. Constitutive expression of Smad3 in the presence of TGF-beta induced apoptotic cell death, with an adverse effect on the cell growth of human lung epithelial cells. Apoptotic cell death could also be induced by forced expression of Smad2 in the presence of TGF-beta, but less efficiently than by that of Smad3. These findings clearly define the distinctions between Smad2 and Smad3 for the first time in that a qualitative difference was observed with regard to the regulation of their expression in response to TGF-beta, while Smad2 and Smad3 appeared to have quantitatively different capabilities regarding the induction of apoptotic cell death in human lung epithelial cells.

• Shou W et al.
• Cardiac defects and altered ryanodine receptor function in mice lacking FKBP12.
• Nature. 1998; 391: 489-92
• Display abstract

• FKBP12, a cis-trans prolyl isomerase that binds the immunosuppressants FK506 and rapamycin, is ubiquitously expressed and interacts with proteins in several intracellular signal transduction systems. Although FKBP12 interacts with the cytoplasmic domains of type I receptors of the transforming growth factor-beta (TGF-beta) superfamily in vitro, the function of FKBP12 in TGF-beta superfamily signalling is controversial. FKBP12 also physically interacts stoichiometrically with multiple intracellular calcium release channels including the tetrameric skeletal muscle ryanodine receptor (RyR1). In contrast, the cardiac ryanodine receptor, RyR2, appears to bind selectively the FKBP12 homologue, FKBP12.6. To define the functions of FKBP12 in vivo, we generated mutant mice deficient in FKBP12 using embryonic stem (ES) cell technology. FKBP12-deficient mice have normal skeletal muscle but have severe dilated cardiomyopathy and ventricular septal defects that mimic a human congenital heart disorder, noncompaction of left ventricular myocardium. About 9% of the mutants exhibit exencephaly secondary to a defect in neural tube closure. Physiological studies demonstrate that FKBP12 is dispensable for TGF-beta-mediated signalling, but modulates the calcium release activity of both skeletal and cardiac ryanodine receptors.

• Letterio JJ, Roberts AB
• TGF-beta: a critical modulator of immune cell function.
• Clin Immunol Immunopathol. 1997; 84: 244-50

• Caussanel V, Tabone E, Hendrick JC, Dacheux F, Benahmed M
• Cellular distribution of transforming growth factor betas 1, 2, and 3 and their types I and II receptors during postnatal development and spermatogenesis in the boar testis.
• Biol Reprod. 1997; 56: 357-67
• Display abstract

• Transforming growth factor betas (TGF betas) 1, 2, and 3 and their types I and II receptors (TGF betas RI and RII) were immunolocalized 1) during testicular development from the perinatal to the adult period and 2) in maturing germ cell populations at successive seminiferous epithelium stages. In the perinatal testis, TGF beta isoforms and receptors were both preponderant in Leydig cells and in spermatogonia. At prepuberty, their expression appeared in Sertoli cells, while germ cells showed specific TGF beta1 and TGF betaRI staining in the spermatocytes. In the adult testis, TGF beta ligands exhibited a preferential tubular distribution. TGF beta1 was mainly detected in young spermatocytes, TGF beta2 in Sertoli cells, and TGF beta3 in Sertoli and premeiotic germ cells. Although the two receptors were systematically observed together in various cells, our data indicate a predominance of one in comparison with the other depending on the cell type. TGF betaRI was predominant in meiotic and differentiated germ cells and TGF betaRII in somatic cells. Finally, in the adult testis, TGF betas 1, 3, and RI showed a germ-cell pattern that depended upon the stage of the seminiferous epithelium cycle. Specifically, staining for the ligands was predominant before meiosis, and TGF betaRI was present particularly during meiosis and spermiogenesis. Together, the temporal and spatial distribution of the TGF beta system components suggests that these signaling molecules may play a crucial role during specific steps of testicular development and during different waves of seminiferous epithelium maturation leading to spermatogenesis.

• Derynck R, Feng XH
• TGF-beta receptor signaling.
• Biochim Biophys Acta. 1997; 1333: 10550-10550

• Dobie K, Cummins P
• Changes in transforming growth factor beta (TGF-beta) expression in the myocardium.
• Biochem Soc Trans. 1997; 25: 443-443

• Bottinger EP, Letterio JJ, Roberts AB
• Biology of TGF-beta in knockout and transgenic mouse models.
• Kidney Int. 1997; 51: 1355-60
• Display abstract

• This paper reviews the basic biology and biochemistry of the TGF-beta isoforms including their unique serine-threonine receptors and signaling intermediates. Dysregulation of TGF-beta expression and/or receptor/signaling function have been implicated in a wide variety of pathologies. We will discuss mechanisms underlying some of these disease processes as gained from study of transgenic mice in which expression of TGF-beta 1 has either been lost by targeted deletion of its gene, is overexpressed in a tissue-specific manner, or blocked by its latency associated peptide.

• Barrack ER
• TGF beta in prostate cancer: a growth inhibitor that can enhance tumorigenicity.
• Prostate. 1997; 31: 61-70
• Display abstract

• A common feature of cancer cells is the autocrine production of growth promoters and the loss of function of tumor suppressors. In our search for such features of prostate cancer, we discovered that transforming growth factor beta 1 (TGF beta 1) levels are higher in prostate cancer than in normal prostate, and prostate cancer cells can activate endogenously-produced latent TGF beta to a bioactive form. Because TGF beta 1 is a potent growth inhibitor of epithelial cells, it seems paradoxical that malignant epithelial cells make high levels of a growth inhibitor. Even prostate cancer cells can be growth-inhibited by TGF beta 1, but only under specific conditions in vitro (plating at low cell density in serum-free medium), and this response is readily disrupted by growth factors, serum, and extracellular matrix, to all of which the cells are exposed in vivo. This explains why prostate cancer cells are resistant to the growth-inhibitory effect of TGF beta in vivo. In vivo, TGF beta 1 actually enhances prostate tumor growth and metastasis, but not by affecting tumor cell proliferation directly. One possibility is that TGF beta affects the host to allow increased numbers of tumor cells to survive and produce progeny. In addition, since prostate cancer cells can still respond to TGF beta, e.g., by increased cell motility, even under conditions that prevent growth inhibition, the ability of TGF beta to enhance tumorigenicity in vivo might also occur via direct effects on the tumor cells themselves. I will discuss new developments in our understanding of TGF beta action, which provide a framework for elucidating the mechanism by which prostate cancer cells have devised a way to protect themselves from being growth-inhibited by TGF beta 1 in vivo. Since the cells retain the ability to be growth-inhibited by TGF beta, indicating that the TGF beta receptors and signaling pathways for growth inhibition are intact, albeit inactive, it might be possible to reactivate this pathway to achieve a therapeutic benefit in vivo.

• Bottinger EP et al.
• Expression of a dominant-negative mutant TGF-beta type II receptor in transgenic mice reveals essential roles for TGF-beta in regulation of growth and differentiation in the exocrine pancreas.
• EMBO J. 1997; 16: 2621-33
• Display abstract

• Using a dominant-negative mutant receptor (DNR) approach in transgenic mice, we have functionally inactivated transforming growth factor-beta (TGF-beta) signaling in select epithelial cells. The dominant-negative mutant type II TGF-beta receptor blocked signaling by all three TGF-beta isoforms in primary hepatocyte and pancreatic acinar cell cultures generated from transgenic mice, as demonstrated by the loss of growth inhibitory and gene induction responses. However, it had no effect on signaling by activin, the closest TGF-beta family member. DNR transgenic mice showed increased proliferation of pancreatic acinar cells and severely perturbed acinar differentiation. These results indicate that TGF-beta negatively controls growth of acinar cells and is essential for the maintenance of a differentiated acinar phenotype in the exocrine pancreas in vivo. In contrast, such abnormalities were not observed in the liver. Additional abnormalities in the pancreas included fibrosis, neoangiogenesis and mild macrophage infiltration, and these were associated with a marked up-regulation of TGF-beta expression in transgenic acinar cells. This transgenic model of targeted functional inactivation of TGF-beta signaling provides insights into mechanisms whereby loss of TGF-beta responsiveness might promote the carcinogenic process, both through direct effects on cell proliferation, and indirectly through up-regulation of TGF-betas with associated paracrine effects on stromal compartments.

• Josso N, di Clemente N
• Serine/threonine kinase receptors and ligands.
• Curr Opin Genet Dev. 1997; 7: 371-7
• Display abstract

• Serine/threonine receptors transduce signals for the TGF-beta family, several members of which, such as decapentaplegic and bone morphogenetic proteins, are involved in early patterning of the embryo. The gene encoding the anti-Mullerian hormone (AMH) receptor has recently been cloned; gene targeting produces the same effects as targeting of the AMH gene itself. Another divergent member of the TGF-beta family, GDNF, signals through Ret, a tyrosine kinase receptor; binding to Ret requires the cooperation of GDNFR-alpha. The signal transduction pathway of serine/threonine receptors is now being intensively studied; the immunophilin FKBP-12 and MAD proteins are known to be involved.

• Heldin CH, Miyazono K, ten Dijke P
• TGF-beta signalling from cell membrane to nucleus through SMAD proteins.
• Nature. 1997; 390: 465-71
• Display abstract

• The recent identification of the SMAD family of signal transducer proteins has unravelled the mechanisms by which transforming growth factor-beta (TGF-beta) signals from the cell membrane to the nucleus. Pathway-restricted SMADs are phosphorylated by specific cell-surface receptors that have serine/threonine kinase activity, then they oligomerize with the common mediator Smad4 and translocate to the nucleus where they direct transcription to effect the cell's response to TGF-beta. Inhibitory SMADs have been identified that block the activation of these pathway-restricted SMADs.

• Miyazono K, Kawabata M
• [TGF-beta signal transduction and Smad]
• Seikagaku. 1997; 69: 424-8

• Yamashita H et al.
• [Functions of the transforming growth factor-beta superfamily in eyes]
• Nippon Ganka Gakkai Zasshi. 1997; 101: 927-47
• Display abstract

• One human body is composed of 6 x 10(13) cells, and eyes are also composed of many cells of different functions. The cellular functions and intercellular interaction are regulated by many regulators including cytokines and growth factors to maintain the homeostasis. The transforming growth factor-beta (TGF-beta) superfamily, a large family of multifunctional factors, regulates various cellular functions, including cellular proliferation, migration, differentiation, apoptosis and extracellular matrix production. The TGF-beta superfamily contains about 30 multifunctional factors, and is divided into several families according to the sequence homology. The TGF-beta family, the activin family, and bone morphogenic proteins belong to the TGF-beta superfamily. TGF-beta superfamily members transduce signals through type I and type II serine/threonine type transmembrane receptors. The signals are transduced from receptors through nuclei by Smad family members, which are phosphorylated by the activated type I receptors and translocate from cytoplasm into nuclei. TGF-beta family members and the TGF-beta superfamily receptor family are expressed in ocular tissues including the cornea, ciliary epithelium, lens epithelium, retina, and blood vessels. This observation suggests the importance of the TGF-beta superfamily in eyes. Smad family members (Smad 1, Smad 2, Smad 3 and Smad 4) are expressed in the cultured retinal pigmant epithelial cell line (D407), in which TGF-beta and activin A stimulate the translocation of Smad 2, but not Smad 1 into nuclei, whereas bone morphogenetic protein (BMP) stimulates that of Smad 1, but not Smad 2. TGF-beta superfamily members play important roles in the pathogenesis of retinal neovascularization and in the wound healing process of corneal tissue. TGF-beta inhibits the endothelial functions, but, stimulates angiogenesis in vivo. TGF-beta is involved in the formation of abnormal connective tissue in corneal wound healing. In these processes, many cytokines and growth factors are involved, interacting with each other and forming networks. It is mandatory to clarify the networks to investigate molecular pathogenesis and new therapeutic agents.

• Pepper MS
• Transforming growth factor-beta: vasculogenesis, angiogenesis, and vessel wall integrity.
• Cytokine Growth Factor Rev. 1997; 8: 21-43
• Display abstract

• Genetic studies have recently revealed a role for transforming growth factor-beta-1 (TGF-beta 1) and its receptors (TGF-beta Rs I and II as well as endoglin) in embryonic vascular assembly and in the establishment and maintenance of vessel wall integrity. The purpose of this review is threefold: first, to reassess previous studies on TGF-beta and endothelium in the light of these recent findings; second, to describe some of the well-established as well as controversial issues concerning TGF-beta and its regulatory role in angiogenesis; and third, to explore the notion of "context' with respect to TGF-beta and endothelial cell function. Although the focus of this review will be on the endothelium, other vascular wall cells are also likely to be important in the pathogenesis of the vascular lesions revealed by genetic studies.

• Whitman M
• Signal transduction. Feedback from inhibitory SMADs.
• Nature. 1997; 389: 549-51

• Baker JC, Harland RM
• From receptor to nucleus: the Smad pathway.
• Curr Opin Genet Dev. 1997; 7: 467-73
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily plays a central role in the specification and patterning of cells in the early embryo. Several years ago, the TGF-beta s were shown to signal through serine/threonine receptor kinases. Now, with the identification of Smad proteins, we can trace the TGF-beta signal transduction pathway from the receptors into the nucleus.

• Hartsough MT, Mulder KM
• Transforming growth factor-beta signaling in epithelial cells.
• Pharmacol Ther. 1997; 75: 21-41
• Display abstract

• Transforming growth factor (TGF)-beta is a potent growth suppressor of epithelial cells. Resistance to TGF-beta, however, occurs frequently in solid tumors of epithelial origin and contributes to the uncontrolled growth of these tumors. Although mutant receptor proteins contribute to TGF-beta insensitivity, deregulation of TGF-beta signaling cascades represents an equally important mechanism underlying TGF-beta resistance. Identification of abnormal regulation of signaling components in tumor epithelial cells will lead to the development of selective therapeutic approaches to repair the relevant signaling cascade(s) and reverse the growth anomaly. Within the past few years, great strides have been made in defining signaling pathways for TGF-beta. For example, our laboratory has demonstrated a direct correlation between TGF-beta-mediated growth inhibition of epithelial cells and activation of Ras and three members of the mitogen-activated protein kinase (MAPK) superfamily. The TGF-beta signaling events were sustained, dose-dependent, and absent in TGF-beta-resistant cells. Further, up-regulation of both p27Kip1 and p21Cip1, nuclear events important for the growth inhibitory effect of TGF-beta, are completely dependent upon the activation of Ras. However, Ras-independent pathways are also activated simultaneously with the Ras/MAPK pathways to mediate the final TGF-beta growth inhibitory outcome. One such pathway includes the SMAD signaling components that control TGF-beta-mediated gene transcription, currently under active study by a number of laboratories, including our own. Future efforts in this field will focus on defining the significance of these signaling proteins and pathways in mediating specific TGF-beta responses. Moreover, additional novel signaling proteins are sure to be identified.

• Alevizopoulos A, Mermod N
• Transforming growth factor-beta: the breaking open of a black box.
• Bioessays. 1997; 19: 581-91
• Display abstract

• Transforming growth factor-beta (TGF-beta) and its related proteins regulate broad aspects of body development, including cell proliferation, differentiation, apoptosis and gene expression, in various organisms. Deregulated TGF-beta function has been causally implicated in the generation of human fibrotic disorders and in tumor progression. Nevertheless, the molecular mechanisms of TGF-beta action remained essentially unknown until recently. Here, we discuss recent progress in our understanding of the mechanism of TGF-beta signal transduction with respect to the regulation of gene expression, the control of cell phenotype and the potential usage of TGF-beta for the treatment of human diseases.

• Gleizes PE et al.
• TGF-beta latency: biological significance and mechanisms of activation.
• Stem Cells. 1997; 15: 190-7
• Display abstract

• Transforming growth factor (TGF-) beta is secreted as a latent complex in which the mature growth factor remains associated with its propeptide. In order to elicit a biological response, the cytokine must be released from the latent complex, a process termed latent TGF-beta activation or TGF-beta formation. Although latent TGF-beta activation is a critical step in the regulation of its activity, little is known about the molecular mechanisms that lead to the production of active TGF-beta. In this article, we present an overview of the data available on this topic, and we propose a tentative model for the mechanism of TGF-beta formation based upon the observations with different cell systems and on recent findings on the structure of the latent TGF-beta complex.

• Jackson SM et al.
• dally, a Drosophila glypican, controls cellular responses to the TGF-beta-related morphogen, Dpp.
• Development. 1997; 124: 4113-20
• Display abstract

• Decapentaplegic (Dpp) is a Drosophila member of the Transforming Growth Factor-beta (TGF-beta)/Bone Morphogenetic Protein (BMP) superfamily of growth factors. Dpp serves as a classical morphogen, where concentration gradients of this secreted factor control patterning over many cell dimensions. Regulating the level of Dpp signaling is therefore critical to its function during development. One type of molecule proposed to modulate growth factor signaling at the cell surface are integral membrane proteoglycans. We show here that division abnormally delayed (dally), a Drosophila member of the glypican family of integral membrane proteoglycans is required for normal Dpp signaling during development, affecting cellular responses to this morphogen. Ectopic expression of dally+ can alter the patterning activity of Dpp, suggesting a role for dally+ in modulating Dpp signaling strength. These findings support a role for members of the glypican family in controlling TGF-beta/BMP activity in vivo by affecting signaling at the cell surface.

• Miyazono K
• TGF-beta receptors and signal transduction.
• Int J Hematol. 1997; 65: 97-104
• Display abstract

• TGF-beta inhibits the growth of many cell types, including hematopoietic cells and lymphocytes. TGF-beta transduces signals through two different types of serine/threonine kinase receptors, type I (T beta R-I) and type II (T beta R-II). T beta R-II is a primary binding protein for the ligands, and T beta R-I is an effector protein, which determines the specificity of signals. Type III receptor (betaglycan) and endoglin play more indirect roles; i.e. delivery of ligands to the signaling receptors. Various molecules, including farnesyl transferase-alpha, Mothers against dpp (Mad)-related proteins, and a novel MAPKKK (TAK1), have been suggested to participate in the signal transduction of TGF-beta receptors. TGF-beta receptors and Mad-related proteins have been found to act as tumor suppressor genes in various tumors, including colorectal cancers and T-cell lymphoma.

• Nakayama M et al.
• Roles of TGF-beta and latent TGF-beta-binding protein in glomerulosclerosis induced by two consecutive injections of monoclonal antibody 1-22-3 in rats.
• Nephron. 1997; 76: 82-9
• Display abstract

• The present study demonstrated the elevated synthesis and gene expressions of transforming growth factor beta (TGF-beta) or latent TGF-beta binding protein (LTBP) in an irreversible glomerulosclerosis rat model induced by two consecutive injections of monoclonal antibody (MoAb) 1-22-3. The rats were intravenously injected with 500 microg of MoAb 1-22-3 either once or twice at an interval of 2 weeks. The rats were sacrificed at 24 h, 1 week, 2 weeks or 16 weeks after the last injection. At 24 h, the mesangiolytic changes in the rats with two injections of MoAb 1-22-3 were similar to those in the rats with one injection. The glomerular matrix score in the rats with two injections was significantly higher than that in the rats with one injection at weeks 1, 2 or 16. An increased LTBP localization in the glomeruli of the rats at week 1 after either one or two injections was detected in the segmentally expanded mesangial matrix. Moreover, LTBP in the glomeruli of rats at week 1 after two injections appeared to be more strongly stained in the enlarged mesangial matrix than that in the rats after one injection. A TGF-beta bioassay using mink lung epithelial cells revealed that the total TGF-beta in the glomerular culture conditioned medium in the rats at week 1 after two injections was significantly larger than that in the rats after one injection. A Northern blotting analysis of the glomeruli showed that both the expressions of TGF-beta and LTBP mRNA in the rats after two injections were higher than those in the rats after one injection. These findings suggested that the elevated TGF-beta or LTBP may thus be related to the irreversible glomerulosclerosis that was induced by two injections of MoAb 1-22-3 into rats.

• Franklin TJ
• Therapeutic approaches to organ fibrosis.
• Int J Biochem Cell Biol. 1997; 29: 79-89
• Display abstract

• Scarring of superficial tissues and chronic fibroses of major organs present major medical problems ranging from disfigurement to progressive disability and death. Growing understanding of the cellular and molecular events, which are common to these intractable disorders, now provides a favourable basis for the development of effective drug therapies. Much attention is focussed on the roles of the many cytokines and growth factors, which contribute to the fibrogenic process. The transforming growth factor (TGF)-beta 1 and 2 isoforms are among the most significant of these and approaches to control their activity include blocking the activation of latent TGF-beta, preventing the ligand-receptor interactions and the inhibition of down-stream signal transduction. Concerns regarding possible risks of the long-term suppression of TGF-beta function point to connective tissue growth factor (CTGF) as a possible alternative target. CTGF is induced by and appears to mediate at least some of the fibrogenic actions of TGF-beta, although not its important antimitogenic activity on epithelial cells. The fibrogenic effects of endothelins and angiotensin II have aroused considerable interest in the anti-fibrotic potential of antihypertensive agents designed primarily to limit the vasoconstrictive activities of these peptides. Polypeptides including interferons alpha and gamma, relaxin, TGF-beta 3 and hepatocyte growth factor, all show an ability to limit fibrogenesis in either clinical or experimental situations. Finally, inhibitors of the enzymes required for the post-translational processing of collagens, including prolyl 4-hydroxylase, C-proteinase and lysyl oxidase provide a more direct means of reducing the deposition of fibrillar collagens into the extracellular matrix although the potentially adverse effects of sustained manipulation of collagen metabolism remain to be investigated.

• Yuasa Y
• [TGF-beta and its receptors]
• Gan To Kagaku Ryoho. 1997; 24: 1460-4
• Display abstract

• TGF-beta is a member of a superfamily associated with development and regulation of cell growth. Recently, its two receptors and intracellular components of signal transduction, a Smad family, have been identified. The type II receptor gene has been found to be mutated specifically in hereditary colon cancers, HNPCC. Alterations of the Smad 4 gene were also detected in pancreatic and colorectal cancers.

• Nakao A et al.
• Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling.
• Nature. 1997; 389: 631-5
• Display abstract

• TGF-beta signals from the membrane to the nucleus through serine/threonine kinase receptors and their downstream effectors, termed SMAD proteins. The activated TGF-beta receptor induces phosphorylation of two such proteins, Smad2 and Smad3, which form hetero-oligomeric complex(es) with Smad4/DPC4 that translocate to the nucleus, where they then regulate transcriptional responses. However, the mechanisms by which the intracellular signals of TGF-beta are switched off are unclear. Here we report the identification of Smad7, which is related to Smad6. Transfection of Smad7 blocks responses mediated by TGF-beta in mammalian cells, and injection of Smad7 RNA into Xenopus embryos blocks activin/TGF-beta signalling. Smad7 associates stably with the TGF-beta receptor complex, but is not phosphorylated upon TGF-beta stimulation. TGFbeta-mediated phosphorylation of Smad2 and Smad3 is inhibited by Smad7, indicating that the antagonistic effect of Smad7 is exerted at this important regulatory step. TGF-beta rapidly induces expression of Smad7 mRNA, suggesting that Smad7 may participate in a negative feedback loop to control TGF-beta responses.

• Imamura T et al.
• Smad6 inhibits signalling by the TGF-beta superfamily.
• Nature. 1997; 389: 622-6
• Display abstract

• SMAD proteins have been identified as signalling mediators of the TGF-beta superfamily, which is involved in a range of biological activities including cell growth, morphogenesis, development and immune responses. Smad1, Smad2, Smad3 and Smad5 are ligand-specific: Smadl and Smad5 transduce signals from bone morphogenetic proteins, and Smad2 and Smad3 mediate signalling by TGF-beta and activin, whereas Smad4 acts as a common signalling component. For example, Smad2 is phosphorylated by the TGF-beta type I receptor upon ligand binding, forms a heteromer with Smad4, and then translocates into the nucleus where it activates transcription. Here we report the isolation of Smad6 in the mouse. Smad6 is quite different in structure from the other SMAD proteins, and forms stable associations with type I receptors. Smad6 interferes with the phosphorylation of Smad2 and the subsequent heteromerization with Smad4, but does not inhibit the activity of Smad3. Smad6 also inhibits the phosphorylation of Smad1 that is induced by the bone morphogenetic protein type IB receptor. These data indicate that signals of the TGF-beta superfamily are regulated both positively and negatively by members of the SMAD family.

• Jurgensmeier JM, Panse J, Schafer R, Bauer G
• Reactive oxygen species as mediators of the transformed phenotype.
• Int J Cancer. 1997; 70: 587-9
• Display abstract

• Reactive oxygen species (ROS) are known to be involved in different pro- and anticarcinogenic mechanisms. However, their influence on the maintenance of the transformed phenotype has not been studied so far. Here we show that the anchorage-independent growth of transformed murine fibroblasts is inhibited by antioxidants and radical scavengers in a concentration-dependent and reversible manner. These agents also reduce TGF-beta-dependent stimulation of colony formation in soft agar, pointing to their specific interference with TGF-beta-triggered signal chains involved in the maintenance of the transformed state.

• Kojima S
• [Regulation of tissue-associated fibrinolysis by retinoids: mechanism and consequence of retinoid-PA-TGF-beta system]
• Seikagaku. 1997; 69: 104-8

• Claudepierre P et al.
• A relationship between TGF-beta 1 or IL-6 plasma levels and clinical features of spondyloarthropathies.
• Br J Rheumatol. 1997; 36: 400-1

• Nakao A et al.
• Identification of Smad2, a human Mad-related protein in the transforming growth factor beta signaling pathway.
• J Biol Chem. 1997; 272: 2896-900
• Display abstract

• Transforming growth factor-beta (TGF-beta) superfamily members are multifunctional cytokines that exert their effects via heteromeric complexes of two distinct serine and threonine kinase receptors. Drosophila mothers against decapentaplegic and related genes in Caenorhabditis elegans, Xenopus, and mammals were shown to function downstream in the intracellular signaling pathways of TGF-beta superfamily members. Here we report the cloning of a Mad-related protein, termed Sma- and Mad-related protein 2 (Smad2). TGF-beta stimulated the phosphorylation and nuclear translocation of Smad2 in nontransfected Mv1Lu cells. In addition, we demonstrated that TGF-beta and activin mediated phosphorylation of Smad2 after its overexpression with appropriate type I and II receptors in COS cells. Smad2 and Smad1 were found to be broadly expressed in human tissues. Smad2 is closely linked to DPC4 on chromosome 18q21.1, a region often deleted in human cancers. Cells that lack Smad2 may escape from TGF-beta-mediated growth inhibition and promote cancer progression.

• McPherron AC, Lawler AM, Lee SJ
• Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member.
• Nature. 1997; 387: 83-90
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily encompasses a large group of growth and differentiation factors playing important roles in regulating embryonic development and in maintaining tissue homeostasis in adult animals. Using degenerate polymerase chain reaction, we have identified a new murine TGF-beta family member, growth/differentiation factor-8 (GDF-8), which is expressed specifically in developing and adult skeletal muscle. During early stages of embryogenesis, GDF-8 expression is restricted to the myotome compartment of developing somites. At later stages and in adult animals, GDF-8 is expressed in many different muscles throughout the body. To determine the biological function of GDF-8, we disrupted the GDF-8 gene by gene targeting in mice. GDF-8 null animals are significantly larger than wild-type animals and show a large and widespread increase in skeletal muscle mass. Individual muscles of mutant animals weigh 2-3 times more than those of wild-type animals, and the increase in mass appears to result from a combination of muscle cell hyperplasia and hypertrophy. These results suggest that GDF-8 functions specifically as a negative regulator of skeletal muscle growth.

• Ninomiya-Tsuji J, Matsumoto K, Shibuya H
• [TGF-beta signaling]
• Tanpakushitsu Kakusan Koso. 1997; 42: 1517-24

• Deng S et al.
• Transfer of genes for IL-10 and TGF-beta to isolated human pancreatic islets.
• Transplant Proc. 1997; 29: 2206-2206

• Singer MA, Penton A, Twombly V, Hoffmann FM, Gelbart WM
• Signaling through both type I DPP receptors is required for anterior-posterior patterning of the entire Drosophila wing.
• Development. 1997; 124: 79-89
• Display abstract

• The imaginal disk expression of the TGF-beta superfamily member DPP in a narrow stripe of cells along the anterior-posterior compartment boundary is essential for proper growth and patterning of the Drosophila appendages. We examine DPP receptor function to understand how this localized DPP expression produces its global effects upon appendage development. Clones of saxophone (sax) or thick veins (tkv) mutant cells, defective in one of the two type I receptors for DPP, show shifts in cell fate along the anterior-posterior axis. In the adult wing, clones that are homozygous for a null allele of sax or a hypomorphic allele of tkv show shifts to more anterior fates when the clone is in the anterior compartment and to more posterior fates when the clone is in the posterior compartment. The effect of these clones upon the expression pattern of the downstream gene spalt-major also correlates with these specific shifts in cell fate. The similar effects of sax null and tkv hypomorphic clones indicate that the primary difference in the function of these two receptors during wing patterning is that TKV transmits more of the DPP signal than does SAX. Our results are consistent with a model in which a gradient of DPP reaches all cells in the developing wing blade to direct anterior-posterior pattern.

• Reiss M
• Transforming growth factor-beta and cancer: a love-hate relationship?
• Oncol Res. 1997; 9: 447-57
• Display abstract

• This commentary proposes the working hypothesis that transforming growth factor-beta (TGF-beta) can play two different and opposite roles with respect to the process of malignant progression. During the earliest stages of carcinogenesis, TGF-beta can act as potent tumor suppressor and may mediate the actions of chemopreventive agents such as retinoids and tamoxifen. However, at some point during the development and progression of malignant neoplasms, bioactive TGF-betas make their appearance in the tumor microenvironment and the tumor cells appear to escape from TGF-beta-dependent growth arrest. Evidence is accumulating rapidly that this TGF-beta resistance is the consequence of inactivating mutations in any one of the genes that encode signaling intermediates, which include the type I and type II TGF-beta receptors, as well as several Smad proteins. The potential implications of the phenotypic switch from TGF-beta sensitive to TGF-beta resistance that occurs during carcinogenesis for cancer prevention and treatment are discussed.

• Bonyadi M et al.
• Mapping of a major genetic modifier of embryonic lethality in TGF beta 1 knockout mice.
• Nat Genet. 1997; 15: 207-11
• Display abstract

• The transforming growth factor beta 1 (TGF beta 1) signalling pathway is important in embryogenesis and has been implicated in hereditary haemorrhagic telangiectasia (HHT), atherosclerosis, tumorigenesis and immunomodulation. Therefore, identification of factors which modulate TGF beta 1 bioactivity in vivo is important. On a mixed genetic background, approximately 50% Tgfb1-/- conceptuses die midgestation from defective yolk sac vasculogenesis. The other half are developmentally normal but die three weeks postpartum. Intriguingly, the vascular defects of Tgfb1-/- mice share histological similarities to lesions seen in HHT patients. It has been suggested that dichotomy in Tgfb1-/- lethal phenotypes is due to maternal TGF beta 1 rescue of some, but not all, Tgfb1-/- embryos12. Here we show that the Tgfb1-/- phenotype depends on the genetic background of the conceptus. In NIH/Ola, C57BL/6J/Ola and F1 conceptuses, Tgfb1-/- lethality can be categorized into three developmental classes. A major codominant modifier gene of embryo lethality was mapped to proximal mouse chromosome 5, using a genome scan for non-mendelian distribution of alleles in Tgfb1-/- neonatal animals which survive prenatal lethality. This gene accounts for around three quarters of the genetic effect between mouse strains and can, in part, explain the distribution of the three lethal phenotypes. This approach, using neonatal DNA samples, is generally applicable to identification of loci that influence the effect of early embryonic lethal mutations, thus furthering knowledge of genetic interactions that occur during early mammalian development in vivo.

• Patterson GI, Koweek A, Wong A, Liu Y, Ruvkun G
• The DAF-3 Smad protein antagonizes TGF-beta-related receptor signaling in the Caenorhabditis elegans dauer pathway.
• Genes Dev. 1997; 11: 2679-90
• Display abstract

• Signals from TGF-beta superfamily receptors are transduced to the nucleus by Smad proteins, which transcriptionally activate target genes. In Caenorhabditis elegans, defects in a TGF-beta-related pathway cause a reversible developmental arrest and metabolic shift at the dauer larval stage. Null mutations in daf-3 suppress mutations in genes encoding this TGF-beta signal, its receptors, and associated Smad signal transduction proteins. daf-3 encodes a Smad protein that is most closely related to mammalian DPC4, and is expressed throughout development in many of the tissues that are remodeled during dauer development. DAF-4, the type II TGF-beta receptor in this pathway, is also expressed in remodeled tissues. These data suggest that the DAF-7 signal from sensory neurons acts as a neuroendocrine signal throughout the body to directly regulate developmental and metabolic shifts in tissues that are remodeled during dauer formation. A full-length functional DAF-3/GFP fusion protein is predominantly cytoplasmic, and this localization is independent of activity of the upstream TGF-beta-related pathway. However, this fusion protein is associated with chromosomes in mitotic cells, suggesting that DAF-3 binds DNA directly or indirectly. DAF-3 transgenes also interfere with dauer formation, perhaps attributable to a dosage effect. A truncated DAF-3/GFP fusion protein that is predominantly nuclear interferes with dauer formation, implying a role for DAF-3 in the nucleus. These data suggest that DAF-7 signal transduction antagonizes or modifies DAF-3 Smad activity in the nucleus to induce reproductive development; when DAF-7 signals are disabled, unmodified DAF-3 Smad activity mediates dauer arrest and its associated metabolic shift. Therefore, daf-3 is unique in that it is antagonized, rather than activated, by a TGF-beta pathway.

• Kulkarni AB, Karlsson S
• Inflammation and TGF beta 1: lessons from the TGF beta 1 null mouse.
• Res Immunol. 1997; 148: 453-6

• Das SK, Lim H, Wang J, Paria BC, BazDresch M, Dey SK
• Inappropriate expression of human transforming growth factor (TGF)-alpha in the uterus of transgenic mouse causes downregulation of TGF-beta receptors and delays the blastocyst-attachment reaction.
• J Mol Endocrinol. 1997; 18: 243-57
• Display abstract

• In the mouse, the initiation of the attachment reaction between the blastocyst trophectoderm and luminal epithelium of the receptive uterus occurs in the evening (2200-2300 h) of day 4 of pregnancy (day 1 = vaginal plug) and is followed by proliferation and differentiation of stromal cells into decidual cells at the sites of blastocyst attachment. This investigation demonstrates that an inappropriate expression of the human transforming growth factor alpha (hTGF-alpha) transgene in the uterus under the direction of a mouse metallothionein-I promoter downregulates uterine expression of TGF-beta receptor subtypes and delays the initiation of implantation (attachment reaction) resulting in delayed parturition. This delay in the attachment reaction is accompanied by deferred uterine expression of amphiregulin. The results suggest that a coordinated 'cross-talk' between the signaling pathways executed by epidermal growth factor-like growth factors and TGF-beta 5 is important for the normal implantation process.

• Ramp U, Jaquet K, Reinecke P, Nitsch T, Gabbert HE, Gerharz CD
• Acquisition of TGF-beta 1 resistance: an important progression factor in human renal cell carcinoma.
• Lab Invest. 1997; 76: 739-49
• Display abstract

• The balance of growth regulation in tumors can be profoundly disturbed by defects in the transforming growth factor-beta 1 (TGF-beta 1) system. Thus far, investigations into the secretion of TGF-beta 1, the expression of its type I, II, and III receptors, as well as the functional intactness of the TGF-beta 1 signal transduction pathways in human renal cell carcinomas (RCC) have been lacking. The objective of the present study, therefore, was to elucidate the role of the TGF-beta 1 system in RCC. We were able to determine the status of this system in 20 primary RCC and in 30 newly established human RCC cell lines of all major histologic types. All primary RCC showed expression of TGF-beta 1 and its type I and II receptors by immunohistochemistry, irrespective of histologic type. In vitro, all RCC cell lines secreted TGF-beta 1 protein as a biologically inactive complex, which cannot interact with cell-surface receptors. Type I ALK-5-receptor mRNA and protein were detected in 29 RCC cell lines, whereas type I ALK-2-receptor mRNA was found in all cell lines. Type II-receptor mRNA and protein could be demonstrated in all cell lines analyzed, and type III-receptor mRNA was observed in five RCC cell lines. Exogenously added, biologically active TGF-beta 1 (1 ng/ml) resulted in significant (p < 0.05) inhibition of proliferation in 14 of 30 RCC cell lines. Sixteen of thirty RCC cell lines, however, proved to be TGF-beta 1-resistant. This resistance could not be explained by mutations in two "hot spot" regions of the type II-receptor gene (bp 622 to 795 and bp 1868 to 2019), as was demonstrated by DNA sequencing in the TGF-beta 1-resistant RCC cell lines. In conclusion, our observations are the first to provide evidence of an "escape" from the negative growth control of TGF-beta 1 by a significant proportion of RCC, suggesting that the acquisition of TGF-beta 1 resistance is an important progression factor for human RCC.

• Markowitz SD
• Atherosclerosis, just another cancer?
• J Clin Invest. 1997; 100: 2143-5

• Hayashi H et al.
• The MAD-related protein Smad7 associates with the TGFbeta receptor and functions as an antagonist of TGFbeta signaling.
• Cell. 1997; 89: 1165-73
• Display abstract

• TGFbeta signaling is initiated when the type I receptor phosphorylates the MAD-related protein, Smad2, on C-terminal serine residues. This leads to Smad2 association with Smad4, translocation to the nucleus, and regulation of transcriptional responses. Here we demonstrate that Smad7 is an inhibitor of TGFbeta signaling. Smad7 prevents TGFbeta-dependent formation of Smad2/Smad4 complexes and inhibits the nuclear accumulation of Smad2. Smad7 interacts stably with the activated TGFbeta type I receptor, thereby blocking the association, phosphorylation, and activation of Smad2. Furthermore, mutations in Smad7 that interfere with receptor binding disrupt its inhibitory activity. These studies thus define a novel function for MAD-related proteins as intracellular antagonists of the type I kinase domain of TGFbeta family receptors.

• Zhang F, Endo S, Cleary LJ, Eskin A, Byrne JH
• Role of transforming growth factor-beta in long-term synaptic facilitation in Aplysia.
• Science. 1997; 275: 1318-20
• Display abstract

• The role of transforming growth factor-beta (TGF-beta) in long-term synaptic facilitation was examined in isolated Aplysia ganglia. Treatment with TGF-beta1 induced long-term facilitation (24 and 48 hours), but not short-term (5 to 15 minutes) or intermediate-term (2 to 4 hours) facilitation. The long-term effects of TGF-beta1 were not additive with those of serotonin. Moreover, serotonin-induced facilitation was blocked by an inhibitor of TGF-beta. Thus, activation of TGF-beta may be part of the cascade of events underlying long-term sensitization, consistent with the hypothesis that signaling molecules that participate in development also have roles in adult neuronal plasticity.

• Ata AK, Funa K, Olsson Y
• Expression of various TGF-beta isoforms and type I receptor in necrotizing human brain lesions.
• Acta Neuropathol (Berl). 1997; 93: 326-33
• Display abstract

• It is known that transforming growth factor beta (TGF-beta) is involved in the modulation of cell growth, differentiation, and repair following injury. We performed an immunohistochemical study of human brain autopsy and biopsy material for the expression of TGF-beta isoforms beta 1, beta 2 and beta 3, and TGF-beta receptor (T beta R) type I in different cells of necrotizing lesions such as infarction and abscess, and compared them with controls. Various cell types, both inside and in the proximity of lesions, showed immunoreactivity indicating the presence of all three isoforms. Significant values of immunoreaction for various TGF-beta s and T beta R-I were observed in cells such as astrocytes, macrophages, neurons, microvascular endothelial cells, and granulocytes. In the control cases, comprising biopsy material without necrotizing lesions, a prominent TGF-beta 2 immunoreactivity was observed in glial cells and neurons. TGF-beta 1 and TGF-beta 3 reactivity in controls, when compared with TGF-beta 2, was less. T beta R-I antiserum showed clear and distinct signals in the same type of cells as for TGF-beta s in the necrotizing lesions with varying values of significance. Our findings suggest that TGF-beta s and their receptor type I are involved in reactive processes around necrotizing human brain lesions like glial and macrophage responses, angiogenesis, and deposition of extracellular matrix.

• Pintavorn P, Ballermann BJ
• TGF-beta and the endothelium during immune injury.
• Kidney Int. 1997; 51: 1401-12
• Display abstract

• During immune injury, activation of endothelial cells by inflammatory cytokines stimulates leukocyte adhesion to the endothelium, turns the endothelium from an anticoagulant surface to one that is frankly procoagulant, and results in the release of vasoactive mediators and growth factors. Cytokine activation of endothelial cells also results in increased endothelial cell TGF-beta 1 synthesis and enhanced activation of latent TGF-beta, the latter involving a shift of plasmin production from the apical to subendothelial surface. In cytokine-stimulated endothelial cells, TGF-beta hinders leukocyte adhesion and transmigration via inhibition of IL-8 and E-selectin expression. TGF-beta also profoundly diminishes cytokine-stimulated inducible nitric oxide synthase production and instead augments endothelial nitric oxide synthase expression. Thus, some of the TGF-beta actions on endothelium during immune activation can viewed as immunosuppressive. TGF-beta also influences mechanisms of vascular remodeling during the healing phase of immune injury. It stimulates PDGF-B synthesis by endothelial cells, causes bFGF release from subendothelial matrix, and promotes VEGF synthesis by non-endothelial cells. Together these mediators control angiogenesis, a critical component of the vascular repair phenomenon. Further, endothelial cell derived PDGF-B and bFGF influence the proliferation and migration of neighboring cells. Thus, endothelial cells and TGF-beta actions on the endothelium play important roles both during the initial phase of immune injury and during the later remodeling phase.

• Yokoyama-Kobayashi M, Saeki M, Sekine S, Kato S
• Human cDNA encoding a novel TGF-beta superfamily protein highly expressed in placenta.
• J Biochem (Tokyo). 1997; 122: 622-6
• Display abstract

• Recently, we developed a simple method for detecting a secretory signal sequence encoded by a cDNA fragment. In this study, we used this method to select cDNA clones encoding secretory proteins from a human full-length cDNA library. Full-sequencing analysis of the candidate clones revealed that one clone encoded a novel TGF-beta superfamily protein. The clone encodes a protein of 308 amino acids of which the C-terminal region shows a characteristic feature of TGF-beta superfamily proteins: seven conserved cysteine residues at the C-terminal preceded by a putative processing site composed of a basic amino acid repeat. The corresponding transcripts are highly expressed in the placenta, so the novel protein may play an important role in reproduction.

• Glansbeek HL, van der Kraan PM, Lafeber FP, Vitters EL, van den Berg WB
• Species-specific expression of type II TGF-beta receptor isoforms by articular chondrocytes: effect of proteoglycan depletion and aging.
• Cytokine. 1997; 9: 347-51
• Display abstract

• Recently, a new isoform of the type II transforming growth factor beta receptor (TGF-beta RII) was identified. This isoform (TGF-beta RII2) contains an insertion of 25 amino acids in the extracellular domain of the receptor. Using RT-PCR the authors demonstrated that both TGF-beta RII1 and TGF-beta RII2 are expressed by chondrocytes in murine and human articular cartilage. Bovine articular chondrocytes expressed TGF-beta RII1 mRNA but did not express detectable levels of TGF-beta RII2 mRNA, suggesting that the new isoform does not play an important role in normal bovine cartilage physiology. Because TGF-beta responses seem to be age related and differential TGF-beta responses have been described between normal cartilage and cartilage undergoing repair the authors studied if the relative mRNA expression between these isoforms is altered during cartilage repair and aging. No differences in the relative mRNA expression of the two isoforms of the type II TGF-beta receptor could be demonstrated in murine cartilage during aging or during the repair phase after mild PG depletion indicating that it is unlikely that age-related TGF-beta responses and differential TGF-beta responses between normal cartilage and cartilage undergoing repair are the result of differences in the relative expression of the two TGF-beta RII isoforms.

• Dean M
• Towards a unified model of tumor suppression: lessons learned from the human patched gene.
• Biochim Biophys Acta. 1997; 1332: 4352-4352

• Kucich U, Rosenbloom JC, Abrams WR, Bashir MM, Rosenbloom J
• Stabilization of elastin mRNA by TGF-beta: initial characterization of signaling pathway.
• Am J Respir Cell Mol Biol. 1997; 17: 10-6
• Display abstract

• The cytokine transforming growth factor-beta (TGF-beta) has multiple effects on a wide variety of cell types. These effects include modulation of growth and regulation of gene transcription. In a few instances, TGF-beta has also been shown to regulate gene expression posttranscriptionally by altering message stability, but the pathway by which this activity is executed remains largely unknown. In the present work, we demonstrate that TGF-beta 1 has no effect on transcription of the elastin gene in cultured human fetal lung fibroblasts, but does stabilize elastin messenger RNA (mRNA), leading to a dramatic increase in the steady-state level of elastin mRNA. A corresponding increase in production of tropoelastin accompanies the increase in elastin mRNA. Through the use of specific inhibitors, we demonstrate that phosphatidylcholine (PC)-specific phospholipase C (PLC) and protein kinase C (PKC) are involved in mediating the elastin message stabilization. In contrast, G proteins and extracellularly regulated kinases do not appear to be involved. These results suggest that although the TGF-beta signaling pathway leading to message stabilization shares components with that modulating transcription, the message-stabilization pathway also contains diverse other elements.

• Mittl PR, Priestle JP, Cox DA, McMaster G, Cerletti N, Grutter MG
• The crystal structure of TGF-beta 3 and comparison to TGF-beta 2: implications for receptor binding.
• Protein Sci. 1996; 5: 1261-71
• Display abstract

• Transforming growth factors beta belong to a group of cytokines that control cellular proliferation and differentiation. Five isoforms are known that share approximately 75% sequence identity, but exert different biological activities. The structure of TGF-beta 3 was solved by X-ray crystallography and refined to a final R-factor of 17.5% at 2.0 A resolution. Comparison with the structure of TGF-beta 2 (Schlunegger MP, Grutter MG, 1992, Nature 358:430-434; Daopin S, Piez KA, Ogawa Y, Davies DR, 1992, Science 257:369-373) reveals a virtually identical central core. Differences exist in the conformations of the N-terminal alpha-helix and in the beta-sheet loops. In TGF-beta 3, the N-terminal alpha-helix has moved approximately 1 A away from the central core. This movement can be correlated with the mutation of Leu 17 to Val and Ala 47 to Pro in TGF-beta 3. The beta-sheet loops rotate as a rigid body 9 degrees around an axis that runs approximately parallel to the dimer axis. If these differences are recognized by the TGF-beta receptors, they might account for the individual cellular responses. A molecule of the precipitating agent dioxane is bound in a crystal contact, forming a hydrogen bond with Trp 32. This dioxane may occupy a carbohydrate-binding site, because dioxane possesses some structural similarity with a carbohydrate. The dioxane is in contact with two tryptophans, which are often involved in carbohydrate recognition.

• Linkhart TA, Mohan S, Baylink DJ
• Growth factors for bone growth and repair: IGF, TGF beta and BMP.
• Bone. 1996; 19: 112-112
• Display abstract

• Current research is reviewed regarding the actions of three growth factor systems on bone formation: insulin-like growth factors (IGFs), transforming growth factor-betas (TGF betas), and bone morphogenetic proteins (BMPs). Each growth factor family consists of multiple related growth factor genes. TGF betas and BMPs 2-7 are subfamilies of a larger TGF beta superfamily. IGFs, TGF betas and BMPs are produced by osteoblasts and other bone cells and affect osteoblast proliferation and differentiation. They are also incorporated into mineralized bone matrix and retain activity when extracted from bone. Various hormones, growth factors, and mechanical stress influence bone cell production of IGFs, TGF betas, and BMPs. Thus these growth factors may function in local regulation of bone formation. Currently there is much interest in the function of IGF binding proteins, which are also produced by bone cells, in regulating IGF activities in bone. Recently, mechanisms for activation of the TGF beta serine/threonine kinase receptors have been investigated, and receptors for BMPs have been identified which are structurally related to TGF beta receptors. In vivo studies are discussed which demonstrate the applicability of IGFs, TGF betas and BMPs to increasing bone formation systemically, promoting fracture healing and inducing bone growth around implants.

• Reddy KB, Jin G, Karode MC, Harmony JA, Howe PH
• Transforming growth factor beta (TGF beta)-induced nuclear localization of apolipoprotein J/clusterin in epithelial cells.
• Biochemistry. 1996; 35: 6157-63
• Display abstract

• Apolipoprotein J (apoJ)/clusterin was first identified as an 80 kDa secretory glycoprotein present in most body fluids. It has been implicated in a variety of physiological processes including cellular differentiation and apoptosis. We demonstrate here that in addition to the well characterized secreted form of the protein, there exists an intracellular, nuclear form of apoJ. This intracellular form of the protein is induced to accumulate in the nucleus of two epithelial cell lines (HepG2 and CCL64) in response to treatment with transforming growth factor beta (TGF beta). We demonstrate in vitro that apoJ protein can be translated from two in-frame ATG sites. Initiation from the first ATG encodes for the secretory form of apoJ and initiation from the second ATG, located 33 amino acids downstream of the first and lacking the hydrophobic signal sequence, encodes for a truncated apoJ protein. This shorter form of apoJ is not recognized by microsomes and therefore not glycosylated, and we postulate that it is retained intracellularly and targeted to the nucleus due to the presence of an SV40-like nuclear localization sequence (NLS). This mechanism of nuclear targeting of apoJ occurs in cells since the protein isolated from nuclei of TGF beta-treated cells and the in vitro-translated truncated form are identical by V8 protease analysis. These results suggest that the diverse physiological responses attributed to apoJ may be elicited through a common molecular mechanism involving a previously uncharacterized intracellular form of the protein.

• Macias-Silva M, Abdollah S, Hoodless PA, Pirone R, Attisano L, Wrana JL
• MADR2 is a substrate of the TGFbeta receptor and its phosphorylation is required for nuclear accumulation and signaling.
• Cell. 1996; 87: 1215-24
• Display abstract

• MAD-related (MADR) proteins are essential intracellular components of TGFbeta signaling pathways and are regulated by phosphorylation. Here, we demonstrate that MADR2 and not the related protein DPC4 transiently interacts with the TGFbeta receptor and is directly phosphorylated by the complex on C-terminal serines. Interaction of MADR2 with receptors and phosphorylation requires activation of receptor I by receptor II and is mediated by the receptor I kinase. Mutation of the phosphorylation sites generates a dominant negative MADR2 that blocks TGFbeta-dependent transcriptional responses, stably associates with receptors, and fails to accumulate in the nucleus in response to TGFbeta signaling. Thus, transient association and phosphorylation of MADR2 by the TGFbeta receptor is necessary for nuclear accumulation and initiation of signaling.

• Mercier T, Gaillard-Sanchez I, Martel P, Heberden C
• TGF-beta receptors are diminished after retinoid exposure in rat liver epithelial cells.
• J Cell Biochem. 1996; 61: 230-7
• Display abstract

• When rat liver epithelial cells were exposed to retinoic acid or retinol for 24 hr, the levels of transforming growth factor-beta (TGF-beta) receptors were reduced in a dose-dependent way. The decrease appeared after 12 hr of incubation with the retinoids and binding levels remained low until 24 hr after the removal of the molecules. Retinoid treatment induced a fourfold enhancement of transglutaminase (TGase) activity in the cell membranes, and cystamine, an inhibitor of TGase, prevented the decrease of the receptors. Neutralization of TGF-beta by a monoclonal antibody did not suppress the decrease of the binding levels, indicating that decreased TGF-beta binding capacity was not due merely to the internalization of ligand-bound receptors promoted by a stimulation of TCF-beta synthesis. Thus, retinoid treatment resulted in an intense disappearance of the functional receptors from the membranes that seemed to be mediated by increased TGase activity. This phenomenon can represent a strong signal attenuation for TGF-beta following retinoid exposure.

• Lagna G, Hata A, Hemmati-Brivanlou A, Massague J
• Partnership between DPC4 and SMAD proteins in TGF-beta signalling pathways.
• Nature. 1996; 383: 832-6
• Display abstract

• The TGF-beta/activin/BMP superfamily of growth factors signals through heteromeric receptor complexes of type I and type II serine/threonine kinase receptors. The signal originated by TGF-beta-like molecules appears to be transduced by a set of evolutionarily conserved proteins known as SMADs, which upon activation directly translocate to the nucleus where they may activate transcription. Five SMAD proteins have so far been characterized in vertebrates. These factors are related to the mediator of decapentaplegic (dpp) signalling, mothers against dpp (Mad), in Drosophila and to the Sma genes from Caenorhabditis elegans. Smad1 and Smad2 have been shown to mimic the effects of BMP and activin, respectively, both in Xenopus and in mammalian cells, whereas Smad3 (a close homologue of Smad2) and the related protein DPC4, a tumour-suppressor gene product, mediate TGF-beta actions. We report here that DPC4 is essential for the function of Smad1 and Smad2 in pathways that signal mesoderm induction and patterning in Xenopus embryos, as well as antimitogenic and transcriptional responses in breast epithelial cells. DPC4 associates with Smad1 in response to BMP and with Smad2 in response to activin or TGF-beta. DPC4 is therefore a regulated partner of SMADs that function in different signalling pathways of the TGF-beta family.

• Twombly V, Blackman RK, Jin H, Graff JM, Padgett RW, Gelbart WM
• The TGF-beta signaling pathway is essential for Drosophila oogenesis.
• Development. 1996; 122: 1555-65
• Display abstract

• We examine roles of signaling by secreted ligands of the TGF-beta family during Drosophila oogenesis. One family member, the DPP ligand encoded by the decapentaplegic (dpp) gene, is required for patterning of anterior eggshell structures. This requirement presumably reflects the expression pattern of dpp in an anterior subset of somatic follicle cells: the centripetally migrating and the nurse cell-associated follicle cells. Similar requirements are also revealed by mutations in the saxophone (sax)-encoded receptor, consistent with the idea that DPP signaling is, at least in part, mediated by the SAX receptor. A loss of germline sax function results in a block in oogenesis associated with egg chamber degeneration and a failure of the transfer of nurse cell contents to the oocyte, indicating that TGF-beta signaling is required for these events. Some phenotypes of sax mutations during oogenesis suggest that SAX responds to at least one other TGF-beta ligand as well in the posterior follicle cells.

• Chambaz EM, Souchelnitskiy S, Pellerin S, Defaye G, Cochet C, Feige JJ
• Transforming growth factors-beta s: a multifunctional cytokine family. Implication in the regulation of adrenocortical cell endocrine functions.
• Horm Res. 1996; 45: 222-6
• Display abstract

• Knowledge of the structure of the first recognized transforming growth factor-beta (TGF-beta 1) has led to the identification of more than two dozen structurally related peptides which appear of crucial importance in the regulation of cell proliferation, cell differentiation and embryogenesis. TGF-beta 1 and its close homologs (TGF-beta 2-5) are multifunctional peptides whose effects on cell functions are dependent upon the cell type, the environment and the presence of other growth factors. TGF-beta 1 is produced and secreted as a latent macromolecular complex. One of the major steps in the control of TGF-beta activity may thus be its release (activation) from its latent form upon the effect of local factors. Adrenocortical cells may be taken as an example in which autocrine production of TGF-beta may be a component of a negative regulatory loop in balance with the positive effect of a systemic hormone (ACTH) in controlling the expression of the cell steroidogenic differentiated functions. In this system, latent TGF-beta can be activated by an ACTH-induced secreted protein (CISP), a member of the thrombospondin family. This points to the importance of the functional interaction between TGF-beta s and extracellular matrix components in the local regulation of cell activities.

• Azuma M, Tamatani T, Fukui K, Yuki T, Motegi K, Sato M
• Different signalling pathways involved in transforming growth factor-beta 1-induced morphological change and type IV collagen synthesis in simian virus-40-immortalized normal human salivary gland duct and myoepithelial cell clones.
• Arch Oral Biol. 1996; 41: 413-24
• Display abstract

• To understand the specific cell type responsible for the synthesis of basement membrane components of the salivary gland, the effects of transforming growth factor (TGF)-beta 1 on morphological change, cellular proliferation and collagen synthesis were examined in these immortalized duct and myoepithelial cell clones, and the expression forms of their TGF-beta receptors analysed. When TGF-beta 1 was added to the cell clones in vitro, it induced a morphological alteration, with flattening in myoepithelial but not in duct cells. Although the growth of Mv1Lu mink lung epithelial cells was almost completely inhibited by TGF-beta 1, the duct and myoepithelial cells were partially resistant to such inhibition. By immunoblot analysis of immunoprecipitates, p53 was found bound to the simian virus-40 large T antigen, suggesting a functional loss of p53 in regulation of cell-cycle arrest. In the cloned myoepithelial cells but not the duct cells, TGF-beta 1 stimulated the production of type IV collagen. To attempt to understand the distinct responsiveness of cell clones to TGF-beta 1, the expression forms of TGF-beta receptors were examined by affinity cross-linking. Although the intensities of the cross-linked bands of the TGF-beta type II and type III receptors, particularly the type II, were weaker in the duct than the myoepithelial cell clones, the expression of the type II receptor mRNA was consistently detected in both clones. Accordingly, the reduction of TGF-beta 1 binding may have occurred at the post-transcriptional level. These findings imply that the cloned myoepithelial cells but not the cloned duct cells produce type IV collagen in response to TGF-beta 1 through the receptor-mediated signal transduction pathway, which is presumably disrupted in the cloned duct cells.

• Kolodziejczyk SM, Hall BK
• Signal transduction and TGF-beta superfamily receptors.
• Biochem Cell Biol. 1996; 74: 299-314
• Display abstract

• The TGF-beta superfamily includes a large number of related growth and differentiation factors expressed in virtually all phyla. Superfamily members bind to specific cell surface receptors that activate signal transduction mechanisms to elicit their effects. Candidate receptors fall into two primary groups, termed type I and type II receptors. Both types are serine/threonine kinases. Upon activation by the appropriate ligand, type I and type II receptors physically interact to form hetero-oligomers and subsequently activate intracellular signaling cascades, ultimately regulating gene transcription and expression. In addition, TGF-beta binds to a third receptor class, type III, a membrane-anchored proteoglycan lacking the kinase activity typical of signal transducing molecules. Type III receptors appear to regulate ligand availability to type I and type II receptors. Although a number of transduction mechanisms may be available to TGF-beta superfamily members, evidence gathered through the use of specific kinase and G-protein inhibitors and through assays measuring activation and levels of signaling intermediates suggests that at least one signaling pathway interacts with Ras and Raf proteins via a G-protein intermediate. Raf begins the cytoplasmic kinase cascade that leads to gene regulation. The myriad responses regulated by TGF-beta superfamily members makes the understanding of signal transduction mechanisms utilized by these proteins of great interest to a wide range of biological disciplines.

• Lechleider RJ, de Caestecker MP, Dehejia A, Polymeropoulos MH, Roberts AB
• Serine phosphorylation, chromosomal localization, and transforming growth factor-beta signal transduction by human bsp-1.
• J Biol Chem. 1996; 271: 17617-20
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily regulates a multitude of cellular and developmental events. TGF-beta family ligands signal through transmembrane serine/threonine kinase receptors whose downstream effectors are largely unknown. Using genetic data from the fruit fly, we have identified a downstream effector of TGF-beta-induced signaling. TGF-beta signaling protein-1 (BSP-1) is rapidly phosphorylated in response to TGF-beta. Localization of bsp-1 to chromosome 4q28 suggests a role in carcinogenesis. These data suggest that BSP-1 is the prototype of a new class of signaling molecules.

• Serra R, Moses HL
• Tumor suppressor genes in the TGF-beta signaling pathway?
• Nat Med. 1996; 2: 390-1

• Feng XH, Derynck R
• Ligand-independent activation of transforming growth factor (TGF) beta signaling pathways by heteromeric cytoplasmic domains of TGF-beta receptors.
• J Biol Chem. 1996; 271: 13123-9
• Display abstract

• Transforming growth factor beta (TGF-beta) transduces signals through two related serine/threonine kinase receptors, the type I and type II receptors, which have the ability to interact with each other. In the heteromeric complex, the type II receptor is the primary determinant of ligand binding and phosphorylates the cytoplasmic domain of the type I receptor. Using a chimeric receptor strategy, we and others have shown previously that a functional TGF-beta receptor complex requires heteromerization of both extracellular and intracellular domains of type I and type II receptors. In the current study, we show that overexpression of two receptors carrying a heteromeric combination of cytoplasmic domains resulted in ligand-independent responses, further supporting the functional requirement of the two heterologous cytoplasmic domains in TGF-beta signaling. Furthermore, coexpression of only the cytoplasmic domains of both the type I and II receptors or tethering the type II to the type I cytoplasmic domain activated TGF-beta responses in a ligand-independent manner. In cotransfected COS-1 cells, both cytoplasmic domains are associated with each other. Our results indicate that the cytoplasmic domains of the type I and type II TGF-beta receptors physically and functionally interact with each other in the heteromeric complex.

• Lastres P et al.
• Endoglin modulates cellular responses to TGF-beta 1.
• J Cell Biol. 1996; 133: 1109-21
• Display abstract

• Endoglin is a homodimeric membrane glycoprotein which can bind the beta 1 and beta 3 isoforms of transforming growth factor-beta (TGF-beta). We reported previously that endoglin is upregulated during monocyte differentiation. We have now observed that TGF-beta itself can stimulate the expression of endoglin in cultured human monocytes and in the U-937 monocytic line. To study the functional role of endoglin, stable transfectants of U-937 cells were generated which overexpress L- or S- endoglin isoforms, differing in their cytoplasmic domain. Inhibition of cellular proliferation and downregulation of c-myc mRNA which are normally induced by TGF-beta 1 in U-937 cells were totally abrogated in L-endoglin transfectants and much reduced in the S-endoglin transfectants. Inhibition of proliferation by TGF-beta 2 was not altered in the transfectants, in agreement with the isoform specificity of endoglin. Additional responses of U-937 cells to TGF-beta 1, including stimulation of fibronectin synthesis, cellular adhesion, platelet/endothelial cell adhesion molecule 1 (PECAM-1) phosphorylation, and homotypic aggregation were also inhibited in the endoglin transfectants. However, modulation of integrin and PECAM-1 levels and stimulation of mRNA levels for TGF-beta 1 and its receptors R-I, R-II, and betaglycan occurred normally in the endoglin transfectants. No changes in total ligand binding were observed in L-endoglin transfectants relative to mock, while a 1.5-fold increase was seen in S-endoglin transfectants. The degradation rate of the ligand was the same in all transfectants. Elucidating the mechanism by which endoglin modulates several cellular responses to TGF-beta 1 without interfering with ligand binding or degradation should increase our understanding of the complex pathways which mediate the effects of this factor.

• Okuda S, Tamaki K, Ando T, Yanagida T, Fujishima M
• TGF-beta behavior in the progressive process in the focal glomerulosclerosis rat model: the role of latent TGF-beta-binding protein.
• Contrib Nephrol. 1996; 118: 78-85

• Wharton K, Ray RP, Findley SD, Duncan HE, Gelbart WM
• Molecular lesions associated with alleles of decapentaplegic identify residues necessary for TGF-beta/BMP cell signaling in Drosophila melanogaster.
• Genetics. 1996; 142: 493-505
• Display abstract

• We have identified the molecular lesions associated with six point mutations in the Drosophila TGF-beta homologue decapentaplegic (dpp). The sites of these mutations define residues within both the pro and ligand regions that are essential for dpp function in vivo. While all of these mutations affect residues that are highly conserved among TGF-beta superfamily members, the phenotypic consequences of the different alleles are quite distinct. Through an analysis of these mutant phenotypes, both in cuticle preparations and with molecular probes, we have assessed the functional significance of specific residues that are conserved among the different members of the superfamily. In addition, we have tested for conditional genetic interactions between the different alleles. We show that two of the alleles are temperature sensitive for the embryonic functions of dpp, such that these alleles are not only embryonic viable as homozygotes but also partially complement other dpp hypomorphs at low temperatures. Our results are discussed with regard to in vitro mutagenesis data on other TGF-beta-like molecules, as well as with regard to the regulation of dpp cell signaling in Drosophila.

• Zappelli F et al.
• The inhibition of differentiation caused by TGFbeta in fetal myoblasts is dependent upon selective expression of PKCtheta: a possible molecular basis for myoblast diversification during limb histogenesis.
• Dev Biol. 1996; 180: 156-64
• Display abstract

• Embryonic and fetal skeletal myoblasts are responsible for the formation of primary and secondary fibers in mammals, but the mechanism which diversifies their fate is unknown. In vitro, embryonic myoblasts are resistant to the differentiation inhibitory effects of transforming growth factor beta and phorbol esters. Thus, differential expression of specific molecules involved in the transduction of extracellular signals may contribute to the different phenotypes. We report here that protein kinase C theta, but none of the other known protein kinase C isoforms, is selectively expressed in fetal and postnatal muscle cells (at both the myoblast and myotube stage) in vitro and in vivo. By contrast, embryonic myoblasts and myotubes do not express protein kinase C theta in vitro or in vivo. This difference is causally related to a differential response to transforming growth factor beta, since overexpression of protein kinase C theta, but not of protein kinase C alpha or zeta, in embryonic myoblasts makes these cells sensitive to transforming growth factor beta. These data demonstrate for the first time that a protein kinase C isoform is a key component of the signal transduction cascade which follows exposure of myoblasts to transforming growth factor beta. They also suggest a specific role for protein kinase C theta in determining the fate of different myoblasts during muscle histogenesis.

• Brannon M, Kimelman D
• Activation of Siamois by the Wnt pathway.
• Dev Biol. 1996; 180: 344-7
• Display abstract

• The Wnt pathway and the recently described Xenopus homeobox gene Siamois share the ability to mimic Nieuwkoop organizer signaling activity when ectopically expressed, prompting us to ask whether the Wnt pathway can activate Siamois expression. We show that activators of the Wnt pathway, and the dorsalizing substance LiCl, activate Siamois expression in Xenopus animal caps. Furthermore, Siamois expression is shown to be activated by beta-catenin cell-autonomously. Although certain TGF-beta family members, such as Vg1, are potent dorsal mesoderm inducers, the TGF-beta pathway only weakly activated Siamois. Finally, we demonstrate that Siamois expression is eliminated from the marginal zone of UV-irradiated Xenopus embryos, supporting the hypothesis that cortical rotation moves a Wnt-like activity into the dorsal region. These results suggest that formation of the Nieuwkoop organizer is dependent on activation of Siamois by a localized Wnt-like signaling activity, which may act synergistically with a vegetally localized TGF-beta signal.

• Miyazono K
• [Transforming growth factor-beta and its receptors]
• Nippon Yakurigaku Zasshi. 1996; 107: 133-40
• Display abstract

• Transforming growth factor-beta (TGF-beta) is a family of multifunctional proteins that inhibit the growth of most cell types, and these proteins induce the deposition of extracellular matrix. TGF-beta inhibits the growth and migration of endothelial cells in vitro, but induces angiogenesis in vivo. TGF-beta belongs to a larger superfamily known as the TGF-beta superfamily, which includes activins and bone morphogenetic proteins. TGF-beta is produced as latent high molecular weight complexes from producer cells and is then activated by plasmin or thrombospondin. Latent TGF-beta binding protein (LTBP) is a component of the latent TGF-beta complex produced from platelets and many other cell types; LTBP plays an important role for the interaction of the latent TGF-beta complex with extracellular matrix components. TGF-beta binds several cell surface receptors, including type III receptor (betaglycan), endoglin, type II receptor and type I receptor. The type III receptor and endoglin are indirectly involved in the signal transduction. The Type II and type I receptors have intracellular serine/threonine kinase domains. They form a heteromeric complex after ligand binding and are most important for signal transduction; the type II receptor transactivates the type I receptor, which transduces various signals.

• Tio M, Ma C, Moses K
• Extracellular regulators and pattern formation in the developing Drosophila retina.
• Biochem Soc Symp. 1996; 62: 61-75
• Display abstract

• The compound eye of Drosophila melanogaster is composed of about 800 similar facets (or ommatidia) each of which contains 20 cells (8 photoreceptor neurons and 12 accessory cells). The fly's vision depends on the precise geometry of these components, and the developmental systems that produce pattern and cell fate have become models for nervous system development in general. The development of this pattern is progressive and appears to involve several levels of induction and inhibition, mediated by diffusible signals. The analysis of genes identified by mutation has revealed that several diffusible factors act in the early patterning steps of the eye. Genes that encode these factors include hedgehog, decapentaplegic, wingless, spitz and scabrous.

• Nellen D, Burke R, Struhl G, Basler K
• Direct and long-range action of a DPP morphogen gradient.
• Cell. 1996; 85: 357-68
• Display abstract

• During development of the Drosophila wing, the decapentaplegic (dpp) gene is expressed in a stripe of cells along the anteroposterior compartment boundary and gives rise to a secreted protein that exerts a long-range organizing influence on both compartments. Using clones of cells that express DPP, or in which DPP receptor activity has been constitutively activated or abolished, we show that DPP acts directly and at long range on responding cells, rather than by proxy through the short-range induction of other signaling molecules. Further, we show that two genes, optomotor-blind and spalt are transcriptionally activated at different distances from DPP-secreting cells and provide evidence that these genes respond to different threshold concentrations of DPP protein. We propose that DPP acts as a gradient morphogen during wing development.

• Polyak K
• Negative regulation of cell growth by TGF beta.
• Biochim Biophys Acta. 1996; 1242: 185-99

• Nikawa J
• ["Inositol" signal transduction in yeast]
• Tanpakushitsu Kakusan Koso. 1996; 41: 1680-7

• Letourneur O, Goetschy JF, Horisberger M, Grutter MG
• Ligand-induced dimerization of the extracellular domain of the TGF-beta receptor type II.
• Biochem Biophys Res Commun. 1996; 224: 709-16
• Display abstract

• Transforming growth factor-beta (TGF-beta) signals by mediating the association of two distinct transmembrane serine/threonine kinase receptors, the type I (T beta RI) and II (T beta RII). Here, we took advantage of recombinant human T beta RII extracellular domain (T beta RII-ED) to analyze TGF-beta/T beta RII complex formation which is the initial event in the construction of a signaling complex. We found that recombinant T beta RII-ED binds TGF-beta 3 more efficiently than TGF-beta 2 and therefore maintains the native T beta RII binding selectivity for the different TGF-beta isoforms. Biochemical analysis showed that free T beta RII-ED is expressed as a monomer. Upon ligand binding, both TGF-beta 3 and -beta 2 isoforms induce homodimerization of T beta RII-ED, each TGF-beta subunit being able to bind one T beta RII-ED molecule. These results suggested that ligand dependent receptor dimerization may be an important early step in the TGF-beta signaling complex formation.

• Shah NM, Groves AK, Anderson DJ
• Alternative neural crest cell fates are instructively promoted by TGFbeta superfamily members.
• Cell. 1996; 85: 331-43
• Display abstract

• How growth factors influence the fate of multipotent progenitor cells is not well understood. Most hematopoietic growth factors act selectively as survival factors, rather than instructively as lineage determination signals. In the neural crest, neuregulin instructively promotes gliogenesis, but how alternative fates are determined is unclear. We demonstrate that bone morphogenic protein 2 (BMP2) induces the basic-helix-loop-helix protein MASH1 and neurogenesis in neural crest stem cells. In vivo, MASH1+ cells are located near sites of BMP2 mRNA expression. Some smooth muscle differentiation is also observed in BMP2. A related factor, transforming growth factor beta1 (TGFbeta1), exclusively promotes smooth muscle differentiation. Like neuregulin, BMP2 and TGFbeta1 act instructively rather than selectively. The neural crest and hematopoietic systems may therefore utilize growth factors in different ways to generate cellular diversity.

• Massague J, Weis-Garcia F
• Serine/threonine kinase receptors: mediators of transforming growth factor beta family signals.
• Cancer Surv. 1996; 27: 41-64
• Display abstract

• TGF beta represents the largest and most versatile cytokine family known in metazoans. The recent identification of transmembrane serine/threonine kinases as TGF beta family receptors represents a major milestone towards understanding how these factors elicit their varied responses. Genetic and biochemical evidence suggests a general model for the mechanism of activation of these receptors. In this model, the ligand acts as an adaptor, bringing a primary receptor kinase in contact with a second kinase, which becomes phosphorylated and thereby competent to propagate the signal to downstream components. Such a kinase cascade on the membrane defines a new variation in signal transduction. Although many details of this mechanism remain to be clarified, its combinatorial capacity may explain the multifunctional nature of its ligands. Several key genes have been identified whose regulation by these signals mediates cellular responses such as cell cycle arrest. This field is advancing at a fast pace, and the identity of components that carry the receptor signals to these genes should soon be unveiled. It is expected that these advances, as they unfold, will in turn clarify the role of miscreant TGF beta signalling in human diseases, cancer included.

• Saltis J
• TGF-beta: receptors and cell cycle arrest.
• Mol Cell Endocrinol. 1996; 116: 227-32
• Display abstract

• Transforming growth factor-beta (TGF-beta) is the prototypic member of a superfamily of proteins important in normal growth and development. The recent molecular cloning and characterization of TGF-beta receptors represents a major advance in our understanding of the mechanism of action of this cytokine. These studies have revealed that TGF-beta elicits its diverse biological responses following the formation of a heteromeric complex involving two distantly related transmembrane serine/threonine kinases, both of which are required for signalling. Furthermore, there has been very rapid progress in the identification of key components that regulate the cell cycle, including the cyclin-dependent kinases (CDKs) and their partners, the cyclins. Perhaps the most significant development has been the isolation of a family of proteins that bind to and inactivate CDKs. The ability of TGF-beta to regulate the action of these CDK inhibitors results in growth arrest of mammalian cells in G1.

• Bogdanowicz P, Vivien D, Felisaz N, Leon V, Pujol JP
• An inositolphosphate glycan released by TGF-beta mimics the proliferative but not the transcriptional effects of the factor and requires functional receptors.
• Cell Signal. 1996; 8: 503-9
• Display abstract

• Transforming growth factor-beta 1 (TGF-beta 1) is a multifunctional polypeptide that regulates a number of cellular processes including cell growth and deposition of extracellular matrix protein. Despite the fact that the signal transduction by TGF-beta has been intensively studied, the molecular mechanisms of that pathway are not clear. We have studied the possibility that an inositolphosphate glycan (IPG) is involved in transmission of the TGF-beta 1 signal. We show that TGF-beta 1 induces IPG release in both rabbit articular chondrocytes (RAC), which are growth stimulated by the factor and Mv1Lu cell line, which is growth inhibited. This release requires functional TGF-beta heteromeric receptors in these two cell types. We also demonstrate that IPG mimics TGF-beta 1-induced growth stimulation in mesenchymal cells (+100%) and growth inhibition in epithelial cells (-80%). Moreover TGF-beta receptor I (T beta R-I) is not required for inhibition of proliferation induced by IPG since derivated mutants of the Mv1Lu cell line lacking T beta R-I intracellular domain (R-1B) are significantly inhibited (-65%). Additionally, we show that IPG does not take part in the signalling pathway that leads to activation of matrix gene transcription. These results suggest that TGF-beta effects on growth regulation and extracellular matrix synthesis implicate two different signalling pathways, IPG being only involved in growth regulation.

• McPherson JM
• p53/TGF-beta/cancer: an intriguing connection.
• Cytokine Growth Factor Rev. 1996; 7: 295-295

• Attisano L, Wrana JL
• Signal transduction by members of the transforming growth factor-beta superfamily.
• Cytokine Growth Factor Rev. 1996; 7: 327-39
• Display abstract

• Transforming growth factor-beta (TGF beta) superfamily members exert their diverse biological effects through their interaction with heteromeric receptor complexes of transmembrane serine/threonine kinases. Both components of the receptor complex, known as receptor I and receptor II are essential for signal transduction. The composition of these complexes can vary significantly due to the promiscuous nature of the ligands and the receptors, and this diversity of interactions can yield a variety of biological responses. Several receptor interacting proteins and potential mediators of signal transduction have now been identified. Recent advances, particularly in our understanding of the function of Mothers against dpp-related (MADR) proteins, are providing new insights into how the TGF beta superfamily signals its diverse biological activities.

• Ren P, Lim CS, Johnsen R, Albert PS, Pilgrim D, Riddle DL
• Control of C. elegans larval development by neuronal expression of a TGF-beta homolog.
• Science. 1996; 274: 1389-91
• Display abstract

• The Caenorhabditis elegans dauer larva is specialized for dispersal without growth and is formed under conditions of overcrowding and limited food. The daf-7 gene, required for transducing environmental cues that support continuous development with plentiful food, encodes a transforming growth factor-beta (TGF-beta) superfamily member. A daf-7 reporter construct is expressed in the ASI chemosensory neurons. Dauer-inducing pheromone inhibits daf-7 expression and promotes dauer formation, whereas food reactivates daf-7 expression and promotes recovery from the dauer state. When the food/pheromone ratio is high, the level of daf-7 mRNA peaks during the L1 larval stage, when commitment to non-dauer development is made.

• Beddington R
• Left, right, left... turn.
• Nature. 1996; 381: 116-7

• Meno C et al.
• Left-right asymmetric expression of the TGF beta-family member lefty in mouse embryos.
• Nature. 1996; 381: 151-5
• Display abstract

• Examples of lateral asymmetry are often found in vertebrates, such as the heart being on the left side, but the molecular mechanism governing the establishment of this left-right (L-R) handedness is unknown. A diffusible morphogen may determine L-R polarity, but a likely molecule has not so far been identified. Here we report on the gene lefty, a member of the transforming growth factor-beta family, which may encode a morphogen for L-R determination. Lefty protein contains the cysteine-knot motif characteristic of this superfamily and is secreted as a processed form of relative molecular mass 25K-32K. Surprisingly, lefty is expressed in the left half of gastrulating mouse embryos. This asymmetric expression is very transient and occurs just before the first sign of lateral asymmetry appears. In the mouse mutants iv and inv, which cause situs inversus, the sites of lefty expression are inverted, indicating that lefty is downstream of iv and inv. These results suggest that lefty may be involved in setting up L-R asymmetry in the organ systems of mammals.

• Brand T, Schneider MD
• Transforming growth factor-beta signal transduction.
• Circ Res. 1996; 78: 173-9

• Wang T et al.
• The immunophilin FKBP12 functions as a common inhibitor of the TGF beta family type I receptors.
• Cell. 1996; 86: 435-44
• Display abstract

• The immunophilin FKBP12 is an evolutionarily conserved abundant protein; however, its physiological roles remain poorly defined. Here we report that FKBP12 is a common cytoplasmic interactor of TGF beta family type I receptors. FKBP12 binds to ligand-free TGF beta type I receptor, from which it is released upon a ligand-induced, type II receptor mediated phosphorylation of the type I receptor. Blocking FKBP12/type I receptor interaction with FK506 nonfunctional derivatives enhances the ligand activity, indicating that FKBP12 binding is inhibitory to the signaling pathways of the TGF beta family ligands. Overexpression of a myristylated FKBP12 in Mv1Lu cell specifically inhibits two separate pathways activated by TGF beta, and two point mutations on FKBP12 (G89P, I90K) abolish the inhibitory activity of FKBP12, suggesting that FKBP12 may dock a cytoplasmic protein to the type I receptors to inhibit TGF beta family mediated signaling.

• Pujol JP, Galera P, Vivien D, Redini F, Boumediene K
• [Physiopathology of arthrotic cartilage]
• Rev Prat. 1996; 46: 810-810

• Jones CM, Armes N, Smith JC
• Signalling by TGF-beta family members: short-range effects of Xnr-2 and BMP-4 contrast with the long-range effects of activin.
• Curr Biol. 1996; 6: 1468-75
• Display abstract

• BACKGROUND: One way of establishing a morphogen gradient in a developing embryo involves the localized synthesis of an inducing molecule followed by its diffusion into surrounding tissues. The morphogen-like effects of the mesoderm-inducing factor activin provide support for this idea in amphibian development. The questions remain, however, of how activin exerts its long-range effects, and whether long-range signalling is a property of all transforming growth factor beta (TGF-beta) family members. RESULTS: We compare the signalling ranges of activin and two other TGF-beta family members, Xnr-2 and BMP-4. Unlike activin, Xnr-2 and BMP-4 act over short distances. Furthermore, the effects of constitutively active activin receptors are strictly cell-autonomous. These observations suggest that the long-range effects of activin occur through protein diffusion and that "relay' mechanisms are not initiated by any of these TGF-beta family members. Mechanisms limiting the signalling range of Xnr-2 were addressed by studying Xnr-2 processing and secretion. An activin-Xnr-2 fusion protein signals over many cell diameters, suggesting that regulated processing or secretion is one limiting factor. Disaggregation and reaggregation of Xnr-2-producing tissues also extends the range of Xnr-2, suggesting that components of intact tissue restrict spread of the protein. CONCLUSIONS: The long-range effects of activin are likely to occur through the diffusion of activin protein. The short-range effects of Xnr-2 and BMP-4 emphasize that long-range diffusion is not a general property of TGF-beta-related molecules. Finally, signalling ranges may be regulated by constraints on processing or secretion and by interactions with extracellular components of embryonic tissues.

• Belbrahem A, Godden-Kent D, Mittnacht S
• Regulation and activity of the retinoblastoma protein family in growth factor-deprived and TGF(beta)-treated keratinocytes.
• Exp Cell Res. 1996; 225: 286-93
• Display abstract

• The retinoblastoma protein (pRB) and the pRB-related pocket proteins p130 and p107, when bound to DNA via the E2F family of transcription factors, suppress transcription and through this may mediate growth arrest. We show here that in HaCat cells arrested by treatment with TGFbeta the only pocket protein associating with DNA-bound E2F is pRB. This contrasts the situation in HaCat cells arrested via growth factor withdrawal, where we find that both pRB and p130 can bind. The above implies that p130 participates in regulating E2F-dependent genes upon growth factor deprivation but not upon TGFbeta arrest. More importantly perhaps, in TGFbeta-arrested cells pRB alone in association with its partner E2Fs may be in charge and sufficient to control E2F-dependent gene transcription. Although p130 is not associated with a DNA-binding E2F complex in TGFbeta-treated cells, it is present in such cells in its underphosphorylated form. We provide evidence for a serum-induced process that may regulate p130 by a mechanism independent of p130 hyperphosphorylation.

• Brown CB, Boyer AS, Runyan RB, Barnett JV
• Antibodies to the Type II TGFbeta receptor block cell activation and migration during atrioventricular cushion transformation in the heart.
• Dev Biol. 1996; 174: 248-57
• Display abstract

• Epithelial-mesenchymal transformation is a critical event in the development of many organ systems including the heart. Descriptive studies have implicated a number of factors in mediating this transformation, including transforming growth factor beta (TGFbeta). We now report that disruption of a TGFbeta signal transduction complex by antibodies directed against the Type II TGFbeta receptor blocks both the endocardial cell activation and subsequent migration that constitute transformation in the chick atrioventricular (AV) cushion. The Type II receptor was localized to both endothelial and endocardial cells of the chick embryo. Incubation of AV cushion explants from Stage 14, 16, and 18 embryos with antibody resulted in a blockade of AV endocardial cell transformation by greater than 50% as measured by mesenchyme formation. Similarly, the appearance of procollagen Type I, a marker of endocardial cell transformation, was blocked. In addition, within 2 hr after the incubation of activated Stage 18 explants with Type II antibody the rate of migration of transformed cells was decreased by 50%. These data suggest that TGFbeta acts directly on AV cushion endocardial cells to stimulate epithelial-mesenchymal transformation and that TGFbeta mediates at least two distinct components of AV cushion transformation, activation and migration.

• Franchini A, Kletsas D, Ottaviani E
• Immunocytochemical evidence of PDGF- and TGF-beta-like molecules in invertebrate and vertebrate immunocytes: an evolutionary approach.
• Histochem J. 1996; 28: 599-605
• Display abstract

• Immunoreactive platelet-derived growth factor-AB and transforming growth factor-beta 1 were demonstrated in invertebrate and vertebrate immunocytes by an immunocytochemical procedure. These factors are only present in phagocytic cells among invertebrate immunocytes, whereas in vertebrate immunocytes they are found in monocytes, granulocytes, lymphocytes, thrombocytes and platelets. These results, in agreement with previous reports, represent further evidence in favour of the hypothesis that Nature has followed a conservative strategy in using a common pool of signal molecules that have been highly conserved throughout evolution.

• Lawn RM et al.
• Feedback mechanism of focal vascular lesion formation in transgenic apolipoprotein(a) mice.
• J Biol Chem. 1996; 271: 31367-71
• Display abstract

• Apolipoprotein(a) (apo(a)), the distinguishing protein of atherogenic lipoprotein(a), directs accumulation of the lipoprotein(a) particle to sites in the arterial wall where atherosclerotic lipid lesions develop in man and in transgenic mice expressing human apo(a). It has been proposed that focal apo(a) accumulation in the transgenic mouse vessel wall causes the observed severe local inhibition of transforming growth factor-beta (TGF-beta) activity and the consequent activation of the smooth muscle cells, which subsequently accumulate lipid to form lesions if the mice are fed a high fat diet. We show that blocking formation of these vascular lesions by two independent mechanisms, tamoxifen treatment and increasing high density lipoprotein, also abolishes apo(a) accumulation, inhibition of TGF-beta activity, and activation of smooth muscle cells. The data are consistent with a feedback mechanism in which an initial accumulation of apo(a) inhibits local TGF-beta activity, leading to further accumulation of apo(a). Breaking the feedback loop prevents smooth muscle cell activation and therefore lipid lesion development.

• Bonyadi M, Cui W, Nagase H, Akhurst RJ
• The TGF beta type II receptor, Tgfbr2, maps to distal mouse chromosome 9.
• Genomics. 1996; 33: 328-9

• Graff JM, Bansal A, Melton DA
• Xenopus Mad proteins transduce distinct subsets of signals for the TGF beta superfamily.
• Cell. 1996; 85: 479-87
• Display abstract

• Xenopus cDNAs homologous to the Drosophila Mad gene and C. elegans CEM genes have been cloned and functionally analyzed by microinjection into frog embryos. The results show that these genes (Xmad) encode intracellular proteins that act downstream of TGF beta superfamily ligands. Most interesting is the fact that different Xmad proteins produce distinct biological responses. Xmad1 produces ventral mesoderm, apparently transducing a signal for BMP2 and BMP4, whereas Xmad2 induces dorsal mesoderm like Vg1, activin, and nodal. These results suggest that an individual Xmad protein waits poised in the cytoplasm for instruction from a distinct subset of TGF beta ligands and then conveys specific information to the nucleus.

• Wrana JL, Attisano L
• MAD-related proteins in TGF-beta signalling.
• Trends Genet. 1996; 12: 493-6

• Derynck R, Zhang Y
• Intracellular signalling: the mad way to do it.
• Curr Biol. 1996; 6: 1226-9
• Display abstract

• A family of proteins has been identified that transduce a signal inside cells following binding of a TGF-beta family ligand to its cognate receptor. Like the prototype Drosophila protein Mad, many members of the family play key roles in developmental signalling.

• Anders RA, Leof EB
• Chimeric granulocyte/macrophage colony-stimulating factor/transforming growth factor-beta (TGF-beta) receptors define a model system for investigating the role of homomeric and heteromeric receptors in TGF-beta signaling.
• J Biol Chem. 1996; 271: 21758-66
• Display abstract

• Transforming growth factor-beta (TGF-beta) belongs to a family of ligands that regulate cell growth and differentiation. The most commonly observed receptors are referred to as the type I, type II, and type III (betaglycan) TGF-beta receptors. Two receptor models have been presented to account for the various cellular responses to TGF-beta. The first proposes that all TGF-beta signaling results from the formation of a heteromeric type I/type II complex, while the second suggests that distinct type I or type II TGF-beta receptor combinations mediate aspects of TGF-beta signaling. We have addressed this general question relating to TGF-beta signaling by constructing chimeric receptors consisting of the extracellular domain of the granulocyte/macrophage colony-stimulating factor (GM-CSF) alpha or beta receptor fused to the transmembrane and cytoplasmic domain of the type I or type II TGF-beta receptor. Since high affinity GM-CSF binding requires dimerization of the alpha and beta ligand binding subunits, the response elicited by defined type I and/or type II TGF-beta receptor cytoplasmic domain homomers or heteromers can be examined. We show in mesenchymal AKR-2B cells that while TGF-beta-dependent transient luciferase activity, endogenous gene activity, and long-term biological responses are similarly induced by activating the chimeric heteromeric receptors with GM-CSF as the endogenous TGF-beta receptor, chimeric homomeric type I/type I or type II/type II receptors are signaling-incompetent.

• Koli K, Keski-Oja J
• Transforming growth factor-beta system and its regulation by members of the steroid-thyroid hormone superfamily.
• Adv Cancer Res. 1996; 70: 63-94
• Display abstract

• TGF-beta s and their receptors are expressed ubiquitously, and they act as key regulators of many aspects of cell growth, differentiation, and function. Steroid action on target tissues is often associated with increase in TGF-beta isoforms. Regulation of TGF-beta expression and activation is crucial for normal development and growth control. The loss of responsiveness of different tumor cells to the antiproliferative effects of TGF-beta is a common feature in carcinogenesis. Multiple changes are required for the cells to gain complete resistance to TGF-beta growth inhibition (Fynan and Reiss, 1993; Kimchi et al., 1988; Samuel et al., 1992). Although many tumor cells are not growth inhibited by TGF-beta, they respond to TGF-beta treatment by changes in the expression of matrix components and enhanced proteolytic activity (KeskiOja et al., 1988). Agents that induce TGF-beta production in target tissues can have a chemopreventive or chemotherapeutic value for the management of epithelial malignancies. Conversely, data supporting a positive role for TGF-beta in established tumor progression are beginning to emerge (Arteaga et al., 1993a,b; Barrett-Lee et al., 1990; Arrick et al., 1992 ; E. A. Thompson et al., 1991). In later stages of tumor development, cell proliferation is often not inhibited by TGF-beta, and tumor cells secrete large amounts of this growth factor (Fynan and Reiss, 1993). In vivo TGF-beta secreted by tumor or stromal cells can influence host responses such as a natural killer cell function and thus indirctly support tumor cell viability (Arteaga et al., 1993b). TGF-beta may also affect tumor growth indirectly by stromal effects and promotion of angiogenesis. TGF-beta may also be involved in the progression of breast tumors from the steroid-sensitive to steroid-insensitive state (King et al., 1989). Understanding of the net effect of TGF-beta in different stages of tumor development is critical for the evaluation of its therapeutic value in cancer treatment.

• Hill CS
• Signalling to the nucleus by members of the transforming growth factor-beta (TGF-beta) superfamily.
• Cell Signal. 1996; 8: 533-44

• Tomoda T et al.
• Transforming growth factor-beta is a survival factor for neonate cortical neurons: coincident expression of type I receptors in developing cerebral cortices.
• Dev Biol. 1996; 179: 79-90
• Display abstract

• Transforming growth factor-beta (TGF-beta) is a multifunctional polypeptide which plays a crucial role in the regulation of cell proliferation, differentiation, and organogenesis. In the present study, we investigated the expression of signaling receptors for TGF-beta in developing mice by in situ hybridization, revealing a significant difference in the expression of TGF-beta type I and type II receptors. Unexpectedly, the TGF-beta type I receptors were exclusively expressed without any detectable expression of the TGF-beta type II receptors in developing cerebral cortices. In primary cortical neurons, a neutralizing antibody for TGF-beta significantly reduced the expression of bcl-2 and subsequently induced neuronal cell death, indicating that TGF-beta functions as a survival factor for cortical neurons in vitro. Consistent with the result of in situ hybridization, the TGF-beta, type I but not type II receptors were detected in primary cortical neurons by affinity crosslink and RT-PCR analyses. The concomitant expression of TGF-beta2 and the TGF-beta type I receptors in developing cerebral cortices suggests that the TGF-beta signaling system plays a pivotal role in neuronal differentiation and that unidentified components may be involved in TGF-beta signaling in the development of the central nervous system.

• Oshima M, Oshima H, Taketo MM
• TGF-beta receptor type II deficiency results in defects of yolk sac hematopoiesis and vasculogenesis.
• Dev Biol. 1996; 179: 297-302
• Display abstract

• TGF-beta signaling is mediated through two types of serine/threonin kinase-containing receptors, type I (TGF-betaRI) and type II (TGF-betaRII), which form a heteromeric complex. In this signaling complex, ligand binding TGF-betaRII phosphorylates and thereby activates the TGF-betaRI to signal downstream pathways. To determine the role of TGF-betaRII in embryogenesis, we have generated a TGF-betaRII gene (Tgfbr2) knockout mouse line. The heterozygous Tgfbr2 knockout mice are developmentally normal. The homozygous Tgfbr2 mutation causes defects in the yolk sac hematopoiesis and vasculogenesis, resulting in an embryonic lethality around 10.5 days of gestation. This phenotype is indistinguishable from the previously reported embryonic lethality by the homozygous TGF-beta1 gene (Tgfb1) null mutation. In addition, we have generated chimeric mice using a Tgfbr2 (-/-) embryonic stem cell line. Some chimeric mice showed several types of congenital anomalies, suggesting that TGF-beta II is important for normal development in a variety of organs.

• Dean M
• Polarity, proliferation and the hedgehog pathway.
• Nat Genet. 1996; 14: 245-7

• Souchelnytskyi S, ten Dijke P, Miyazono K, Heldin CH
• Phosphorylation of Ser165 in TGF-beta type I receptor modulates TGF-beta1-induced cellular responses.
• EMBO J. 1996; 15: 6231-40
• Display abstract

• Transforming growth factor-beta (TGF-beta) signals via an oligomeric complex of two serine/threonine kinase receptors denoted TGF-beta type I receptor (TbetaR-I) and type II receptor (TbetaR-II). We investigated the in vivo phosphorylation sites in TbetaR-I and TbetaR-II after complex formation. Phosphorylation of TbetaR-II was observed at residues in the C-terminus (Ser549 and Ser551) and at residues in the juxtamembrane domain (Ser223, Ser226 and Ser227). TGF-beta1 induced in vivo phosphorylation of serine and threonine residues in the juxtamembrane domain of TbetaR-I in a region rich in glycine, serine and threonine residues (GS domain; Thr185, Thr186, Ser187, Ser189 and Ser191), and more N-terminal of this region (Ser165). Phosphorylation in the GS domain has been shown previously to be involved in activation of the TbetaR-I kinase. We show here that phosphorylation of TbetaR-I at Ser165 is involved in modulation of TGF-beta1 signaling. Mutations of Ser165 in TbetaR-I led to an increase in TGF-beta1-mediated growth inhibition and extracellular matrix formation, but, in contrast, to decreased TGF-beta1-induced apoptosis. A transcriptional activation signal was not affected. Mutations of Ser165 changed the phosphorylation pattern of TbetaR-I. These observations suggest that TGF-beta receptor signaling specificity is modulated by phosphorylation of Ser165 of TbetaR-I.

• Jaskoll T, Choy HA, Melnick M
• The glucocorticoid-glucocorticoid receptor signal transduction pathway, transforming growth factor-beta, and embryonic mouse lung development in vivo.
• Pediatr Res. 1996; 39: 749-59
• Display abstract

• Lung morphogenesis has been shown to be regulated by glucocorticoids (CORT). Because CORT has been primarily thought to affect fetal lung development, previous studies have focused on the role of CORT receptor (GR)-mediated regulation of fetal lung development. Although endogenous CORT increases during embryonic and fetal stages and exogenous CORT treatment in vivo and in vitro clearly accelerates embryonic lung development, little is known about the morphoregulatory role of the embryonic CORT-GR signal transduction pathway during lung development. In this study, we characterize the embryonic mouse CORT-GR pathway and demonstrate: stage-specific in situ patterns of GR immunolocalization; similarity in GR relative mobility with progressive (E13 --> E17) development; that embryonic GR can be activated to bind a GR response element (GRE); significantly increasing levels of functional GR with increasing lung maturation; and the presence of heat shock protein (hsp) 70 and hsp90 from early (E13) to late (E17) developmental stages. These results support the purported importance of the embryonic CORT-GR signal transduction pathway in progressive lung differentiation. To demonstrate that the embryonic CORT-GR directed pathway plays a role in lung development, early embryonic (E12) lungs were exposed to CORT in utero and surfactant-associated protein A (SP-A) expression was analyzed; CORT treatment up-regulates SP-A mRNA expression and spatiotemporal protein distribution. Finally, to determine whether CORT-GR-directed pulmonary morphogenesis in vivo involves the modulation of growth factors, we studied the effect of CORT on TGF-beta gene expression. Northern analysis of TGF-beta 1, TGF-beta 2, and TGF-beta 3 transcript levels in vivo indicates that CORT regulates the rate of lung morpho- and histodifferentiation by down-regulating TGF-beta 3 gene expression.

• Ganan Y, Macias D, Duterque-Coquillaud M, Ros MA, Hurle JM
• Role of TGF beta s and BMPs as signals controlling the position of the digits and the areas of interdigital cell death in the developing chick limb autopod.
• Development. 1996; 122: 2349-57
• Display abstract

• The establishment of the digital rays and the interdigital spaces in the developing limb autopod is accompanied by the occurrence of corresponding domains of expression of TGF beta s and BMPs. This study analyzes whether these coincident events are functionally correlated. The experiments consisted of local administration of TGF beta-1, TGF beta-2 or BMP-4 by means of heparin or Affi-gel blue beads to the chick limb autopod in the stages preceding the onset of interdigital cell death. When beads bearing either TGF beta-1 or -2 were implanted in the interdigits, the mesodermal cells were diverted from the death program forming ectopic cartilages or extra digits in a dose- and stage-dependent fashion. This change in the interdigital phenotype was preceded by a precocious ectopic expression of ck-erg gene around the bead accompanied by down-regulation of bmp-4, msx-1 and msx-2 gene expression. When BMP-beads were implanted in the interdigital spaces, programmed cell death and the freeing of the digits were both accelerated. Implantation of beads bearing BMP-4 at the tip of the growing digits was followed by digit bifurcation, accompanied by the formation of an ectopic area of cell death resembling an extra interdigit, both morphologically and molecularly. The death-inducing effect of the BMP beads and the chondrogenic-inducing effect of the TGF beta beads were antagonized by the implantation of an additional bead preabsorbed with FGF-2, which constitutes a signal characteristic of the progress zone. It is concluded that the spatial distribution of digital rays and interdigital spaces might be controlled by a patterned distribution of TGF beta s and BMPs in the mesoderm subjacent to the progress zone.

• Bottner M, Unsicker K, Suter-Crazzolara C
• Expression of TGF-beta type II receptor mRNA in the CNS.
• Neuroreport. 1996; 7: 2903-7
• Display abstract

• Members of the TGF-beta family have been described to bind to a heteromeric complex of two types of serine/threonine kinase receptors, named T beta R-I and T beta R-II. These receptor molecules are essential for TGF-beta-specific signalling. Several type I and type II receptors have been identified by a variety of methods. TGF-beta 2 and 3 are widely expressed in the CNS and exert multiple functions on neurones and glia. Although the T beta R-I molecule is abundant in the CNS, it was unclear whether the type II receptor found in peripheral organs is also expressed in the CNS. Previous, negative findings, seemed to suggest that a novel, as yet undescribed type II receptor may be expressed in the CNS. We used competitive RT-PCR to detect T beta R-II mRNA in rats at different stages of development (E18, P6, adult) and in different tissues. We detected this mRNA in the lung, liver, heart, gut, kidney, and pituitary of adult rats. Surprisingly, and in contrast to previous studies, similar levels of the T beta R-II mRNA were also detected in several regions of the CNS, namely cortex, midbrain, cerebellum, brain stem and hippocampus. We therefore tentatively conclude that TGF-beta 2 and 3 may signal in the brain by means of the same type-I and -II receptors as found in peripheral organs.

• Lawrence DA
• Transforming growth factor-beta: a general review.
• Eur Cytokine Netw. 1996; 7: 363-74
• Display abstract

• Three isoforms of Transforming Growth Factor-beta (TGF-beta 1, beta 2 and beta 3) exist in mammals. They play critical roles in growth regulation and development. Each isoform is encoded by a unique gene on different chromosomes. All three of these growth factors are secreted by most cell types, generally in a latent form, requiring activation before they can exert biological activity. This activation of latent TGF-beta, which may involve plasmin, thrombospondin and possibly acidic microenvironments, appears to be a crucial regulatory step in controlling their effects. The TGF-betas possess three major activities: they inhibit proliferation of most cells, but can stimulate the growth of some mesenchymal cells; they exert immunosuppressive effects; and they enhance the formation of extracellular matrix. Two types of membrane receptors (type I and type II) possessing a serine/threonine kinase activity within their cytoplasmic domains are involved in signal transduction. Inhibition of growth by the TGF-betas stems from a blockage of the cell cycle in late G1 phase. Among the molecular participants concerned in G1-arrest are the Retinoblastoma (Rb) protein and members of the Cyclin/Cyclin-dependent kinase/Cyclin dependent kinase inhibitor families. In the intact organism the TGF-betas are involved in wound repair processes and in starting inflammatory reactions and then in their resolution. The latter effects of the TGF-betas derive in part from their chemotactic attraction of inflammatory cells and of fibroblasts. From gene knockout and from overexpression studies it has been shown that precise regulation of each isoform is essential for survival, at least in the long term. Several clinical applications for certain isoforms have already shown their efficacy and they have been implicated in numerous other pathological situations.

• Tsuiki H, Fukiishi Y, Kishi K
• Relation of TGF-beta 2 to inhibition of limb bud chondrogenesis by retinoid in rats.
• Teratology. 1996; 54: 191-7
• Display abstract

• A newly synthesized retinoid, Am-80 (4-[5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl] carbamoxyl) benzoic acid, induced limb reduction defects in rats in vivo. The present study was designed to investigate the mechanisms of its teratogenic action and also to examine whether transforming growth factor-beta (TGF-beta) is involved in limb reduction defects by using a rat limb bud cell culture system. Am-80 inhibited DNA synthesis and chondrogenesis of limb bud cells from day 12 embryos at concentrations of 1-4 ng/ml in a concentration-dependent manner. Quantities of TGF-beta 1 and 2 in culture medium from limb bud cells treated with Am-80 were evaluated using sandwich ELISAs. Although TGF-beta 1 was not detected in conditioned medium with or without Am-80, TGF-beta 2 increased in a concentration-dependent manner at drug concentrations that inhibited DNA synthesis and chondrogenesis. In addition, when TGF-beta 2 was exogenously added to limb bud cell culture medium, the synthesis of glycosaminoglycan (GAGs) was reduced even without Am-80 treatment. Neutralization of TGF-beta 2 with antibody prevented the inhibition of GAGs synthesis induced by Am-80. These results suggest that TGF-betas, especially beta 2, are involved in the inhibition of chondrogenesis caused by Am-80 in limb bud cells.

• Niehrs C
• Growth factors. Mad connection to the nucleus.
• Nature. 1996; 381: 561-2

• Mishina Y et al.
• Genetic analysis of the Mullerian-inhibiting substance signal transduction pathway in mammalian sexual differentiation.
• Genes Dev. 1996; 10: 2577-87
• Display abstract

• Mullerian-inhibiting substance (MIS) is a member of the transforming growth factor-beta (TGF-beta) gene family. MIS expression in males causes the regression of the Mullerian ducts, an essential process in male sexual differentiation. Recently, an MIS type II receptor gene has been isolated that is expressed during embryogenesis in mesenchymal cells adjacent to the Mullerian duct epithelium and in Sertoli and granulosa cells of the fetal and adult, male and female gonads, respectively. MIS receptor mutant males develop as internal pseudohermaphrodites, possessing a complete male reproductive tract and also a uterus and oviducts, a phenocopy of MIS ligand-deficient male mice. They express both MIS mRNA and protein, showing that ligand was present, but target organs were hormone-insensitive. All produce sperm, but the majority were infertile because the presence of their female reproductive organs blocks sperm transfer into females. Focal seminiferous tubule atrophy accompanied by Leydig cell hyperplasia was observed and began as early as 2 months of age. The phenotype of MIS ligand/MIS receptor double mutant males was indistinguishable from those of each single mutant. MIS receptor/alpha-inhibin double mutant males developed testicular stromal tumors and large fluid-filled uteri that were identical in phenotype to MIS ligand/alpha-inhibin double mutant males. These studies provide in vivo evidence that MIS is the only ligand of the MIS type II receptor, in contrast to the complexity of other TGF-beta gene family signaling pathways.

• Yu K et al.
• The Drosophila decapentaplegic and short gastrulation genes function antagonistically during adult wing vein development.
• Development. 1996; 122: 4033-44
• Display abstract

• TGF-beta-related signaling pathways play diverse roles during vertebrate and invertebrate development. A common mechanism for regulating the activity of TGF-beta family members is inhibition by extracellular antagonists. Recently, the Drosophila short gastrulation (sog) gene was shown to encode a predicted diffusible factor which antagonizes signaling mediated by the TGF-beta-like Decapentaplegic (Dpp) pathway in the early blastoderm embryo. sog and dpp, which are among the earliest zygotic genes to be activated, are expressed in complementary dorsal-ventral domains. The opposing actions of sog and dpp in the early embryo have been highly conserved during evolution as their vertebrate counterparts, chordin and BMP-4, function homologously to define neural versus non-neural ectoderm in Xenopus. Here we exploit the genetically sensitive adult wing vein pattern to investigate the generality of the antagonistic relationship between sog and dpp. We show that dpp is expressed in vein primordia during pupal wing development and functions to promote vein formation. In contrast, sog is expressed in complementary intervein cells and suppresses vein formation. sog and dpp function during the same phenocritical periods (i.e. 16-28 hours after pupariation) to influence the vein versus intervein cell fate choice. The conflicting activities of dpp and sog are also revealed by antagonistic dosage-sensitive interactions between these two genes during vein development. Analysis of vein and intervein marker expression in dpp and sog mutant wings suggests that dpp promotes vein fates indirectly by activating the vein gene rhomboid (rho), and that sog functions by blocking an autoactivating Dpp feedback loop. These data support the view that Sog is a dedicated Dpp antagonist.

• Mann RS, Abu-Shaar M
• Nuclear import of the homeodomain protein extradenticle in response to Wg and Dpp signalling.
• Nature. 1996; 383: 630-3
• Display abstract

• In Drosophila, Decapentaplegic (Dpp) and Wingless (Wg) are two secreted signalling proteins of the transforming growth factor (TGF)-beta and Wnt families, respectively. Although both are often required during development, only a few downstream components of these signalling pathways have been described. Here we present evidence that in the embryonic midgut both signalling pathways control the subcellular localization of the homeodomain protein encoded by the extradenticle (exd) gene. Exd protein is predominantly nuclear in endoderm cells close to Dpp-and Wg-secreting cells of the visceral mesoderm, but is in the cytoplasm in more distant endoderm cells. Both dpp and wg are required for the nuclear localization of Exd in the endoderm, whereas ectopic expression of dpp and wg expands the domain of nuclear Exd. Furthermore, the nuclear import of Exd correlates with the transcription of an exd-dependent reporter gene in the endoderm. Thus one mechanism by which extracellular signals might control pattern is by directing the graded nuclear localization of homeodomain proteins such as Exd that directly control the expression of target genes.

• Burke R, Basler K
• Dpp receptors are autonomously required for cell proliferation in the entire developing Drosophila wing.
• Development. 1996; 122: 2261-9
• Display abstract

• The mammalian growth factor TGFbeta negatively regulates cell proliferation in various systems. Here we provide evidence that another TGFbeta superfamily member, Drosophila Decapentaplegic (Dpp), stimulates cell proliferation. In the developing wing blade, somatic clones lacking the Dpp receptors Punt or Thick veins (Tkv), or lacking Schnurri, a transcription factor involved in Dpp signal interpretation, fail to grow when induced early in larval development. Furthermore the spatial requirement for these signaling components indicates that Dpp has to travel several cell diameters from its source in order to reach all cells that require its signal. The requirement for Tkv also depends on the distance of cells from the source of the Dpp signal. We propose that Dpp can act at a distance to positively control cell proliferation.

• Newfeld SJ, Chartoff EH, Graff JM, Melton DA, Gelbart WM
• Mothers against dpp encodes a conserved cytoplasmic protein required in DPP/TGF-beta responsive cells.
• Development. 1996; 122: 2099-108
• Display abstract

• The proteins necessary for signal transduction in cells responding to ligands of the TGF-beta family are largely unknown. We have previously identified Mad (Mothers against dpp), a gene that interacts with the TGF-beta family member encoded by decapentaplegic (dpp) in Drosophila. Assay of Mad's role in the DPP-dependent events of embryonic midgut development demonstrates that Mad is required for any response of the visceral mesoderm or endoderm to DPP signals from the visceral mesoderm. Replacement of the normal DPP promoter with a heterologous (hsp70) promoter fails to restore DPP-dependent responses in Mad mutant midguts. Experiments utilizing Mad transgenes regulated by tissue-specific promoters show that MAD is required specifically in cells responding to DPP. Immunohistochemical studies localize MAD to the cytoplasm in all tissues examined. Experiments in Xenopus embryos demonstrate that Drosophila MAD can function in the signaling pathway of BMP-4, a vertebrate homolog of dpp. Based on these results, we propose that Mad is a highly conserved and essential element of the DPP signal transduction pathway.

• Wiersdorff V, Lecuit T, Cohen SM, Mlodzik M
• Mad acts downstream of Dpp receptors, revealing a differential requirement for dpp signaling in initiation and propagation of morphogenesis in the Drosophila eye.
• Development. 1996; 122: 2153-62
• Display abstract

• Decapentaplegic (Dpp), a member of the TGF-beta family of cytokines, has been implicated in many patterning processes in Drosophila, including the initial steps of pattern formation in the developing eye. We show that the Mothers against dpp (Mad) gene is required for dpp signaling during eye development. Clonal analysis demonstrates a cell-autonomous function for Mad and genetic interactions indicate that Mad is an essential component of the signal transduction pathway downstream of the Dpp receptors in responding cells. Mad-mediated dpp signaling is absolutely required for the initiation of the morphogenetic furrow in the eye, but has only a minor role in its subsequent propagation across the eye disc. We also present evidence for the repression of wingless transcription by dpp signaling.

• Raftery LA, Wisotzkey RG
• Characterization of Medea, a gene required for maximal function of the Drosophila BMP homolog Decapentaplegic.
• Ann N Y Acad Sci. 1996; 785: 318-20

• Luo K, Lodish HF
• Signaling by chimeric erythropoietin-TGF-beta receptors: homodimerization of the cytoplasmic domain of the type I TGF-beta receptor and heterodimerization with the type II receptor are both required for intracellular signal transduction.
• EMBO J. 1996; 15: 4485-96
• Display abstract

• Transforming growth factor-beta (TGF-beta) affects multiple cellular functions through the type I and type II receptor Ser/Thr kinases (TbetaRI and TbetaRII). Analysis of TGF-beta signaling pathways has been hampered by the lack of cell lines in which both TbetaRI and TbetaRII are deleted, and by the inability to study signal transduction by TbetaRI independently of TbetaRII since TbetaRI does not bind TGF-beta directly. To overcome these problems, we constructed and expressed chimeric receptors with the extracellular domain of the erythropoietin receptor (EpoR) and the cytoplasmic domains of TbetaRI or TbetaRII. When expressed in Ba/F3 cells, which do not express EpoR, Epo induces the formation of a heteromeric complex between cell surface EpoR-TbetaRI and EpoR-TbetaRII chimeras. Neither the EpoR-TbetaRI nor the EpoR-TbetaRII chimera interacts with endogenous TGF-beta receptors. Ba/F3 cells expressing both EpoR-TbetaRI and EpoR-TbetaRII chimeras, but not EpoR-TbetaRI or EpoR-TbetaRII alone, undergo Epo-induced growth arrest. When expressed in Ba/F3 cells in the absence of the EpoR-TbetaRII chimera, EpoR-TbetaRI(T204D), a chimeric receptor with a point mutation in the GS domain of TbetaRI that is autophosphorylated constitutively, triggers growth inhibition in response to Epo. Thus, both homo- and heterodimerization of the cytoplasmic domain of the type I TGF-beta receptor are required for intracellular signal transduction leading to inhibition of cell proliferation. These chimeric receptors provide a unique system to study the function and signal transduction of individual TGF-beta receptor subunits independently of endogenous TGF-beta receptors.

• Munson L, Wilhite A, Boltz VF, Wilkinson JE
• Transforming growth factor beta in bovine placentas.
• Biol Reprod. 1996; 55: 748-55
• Display abstract

• Transforming growth factor beta s (TGF beta) are a family of multifunctional growth factors that are important embryonic morphogens. Because TGF beta s may regulate the development of epitheliochorial placentas, we investigated the location, expression, secretion, and effects of TGF beta s in bovine placentomes and cell cultures derived from chorionic and endometrial epithelia. Placentomes from early second-trimester pregnancies were examined by immunohistochemistry for TGF beta 1, TGF beta 2, and TGF beta 3, and for TGF beta expression in Northern slot-blots. Effects of TGF beta s were assessed in trophoblastic and endometrial epithelial cell lines by DNA synthesis assays. Secretion of TGF beta s by trophoblastic and endometrial epithelial cells was determined using bioassays. All forms of TGF beta were immunolocalized in bovine placentomes. TGF beta mRNA was expressed in chorioallantois, caruncles, and in cultured trophoblastic and endometrial epithelial cells. Endometrial and trophoblastic cells secreted active and latent TGF beta s, and these cells had a transient proliferative response to all forms of TGF beta. These results indicate that TGF beta s are present at the fetal-maternal interface of the bovine placentome and may promote endometrial and chorionic growth.

• Gupta A, Bazer FW, Jaeger LA
• Differential expression of beta transforming growth factors (TGF beta 1, TGF beta 2, and TGF beta 3) and their receptors (type I and type II) in peri-implantation porcine conceptuses.
• Biol Reprod. 1996; 55: 796-802
• Display abstract

• Beta transforming growth factors (TGF beta 1, TGF beta 2, and TGF beta 3) and type I and II TGF beta receptors were immunohistochemically localized in peri-implantation porcine conceptuses (embryos and associated membranes) collected on Day 10 through Day 14 of gestation. Our results indicate specific immunolocalization of TGF beta isoforms and their receptors in conceptuses during these gestational days. In parietal endoderm, TGF beta 1 immunoreactions were weak to undetectable, TGF beta 2 immunoreactions were intense, and TGF beta 3 immunoreactions were intermediate in intensity to TGF beta 2 and TGF beta 1. In contrast to immunoreactions in endoderm, TGF beta 1 and TGF beta 3 immunostaining in trophectoderm (Tr) was intense. Differences in TGF beta 2 immunostaining of Tr were observed from Days 10 to 14 of gestation. A drastic decrease in cytoplasmic immunostaining of ectoderm and mesoderm was detected from Days 12 to 14 for all TGF beta isoforms and type II receptor; however, type I receptor immunoreactions were consistently detected between Days 10 and 14. Concurrent expression of both type I and type II receptors in the peri-implantation conceptuses suggests that porcine conceptuses are capable of binding and responding to TGF beta s during this period. Differential expression of the three TGF beta isoforms suggests different roles for TGF beta s 1, 2, and 3 in conceptus development. Our results suggest possible roles for TGF beta s in early growth and differentiation of the embryo, differentiation of the Tr, and implantation.

• Ferguson EL
• Conservation of dorsal-ventral patterning in arthropods and chordates.
• Curr Opin Genet Dev. 1996; 6: 424-31
• Display abstract

• Dorsal-ventral patterning within the ectodermal and mesodermal germ layers of Drosophila and Xenopus embryos is specified by a system of genes that has been conserved over 500 million years of evolution. In both organisms, the activity of the TGF-beta family member DPP/BMP4 is antagonized by SOG/CHORDIN. A second Xenopus gene, noggin, has a similar biological activity to chordin. Analysis of the action of these genes indicate that Spemann's organizer promotes dorsal cell fates in Xenopus by antagonizing a ventralizing signal encoded by the Bmp4 gene.

• Markowitz SD, Roberts AB
• Tumor suppressor activity of the TGF-beta pathway in human cancers.
• Cytokine Growth Factor Rev. 1996; 7: 93-102
• Display abstract

• The TGF-betas are a family of cytokines with antiproliferative activity on many cell types. Recent findings demonstrate that the TGF-beta receptor complex functions as a tumor suppressor gene in human malignancy. Somatic mutations of the type II subunit of the TGF-beta receptor (RII) have been demonstrated in several different tumor types. RII frameshift mutations within a short coding region polyadenine repeat are particularly characteristic of colon and gastric cancers that also demonstrate the phenotype of microsatellite instability (RER cancers). These and other mutations as in the type I receptor (RI) are associated with both loss of cell surface TGF beta receptors and with resistance of the cancer cells to TGF-beta-induced growth inhibition. Restoration of receptor expression by gene transfection reverses the transformed phenotype in cancer cells that lacked functional RII or RI. These receptors and, by implication, TGF-beta as well as its complete signalling pathway, are thus new and novel additions to the family of human tumor suppressor genes.

• Massague J
• TGFbeta signaling: receptors, transducers, and Mad proteins.
• Cell. 1996; 85: 947-50

• Savage C et al.
• Caenorhabditis elegans genes sma-2, sma-3, and sma-4 define a conserved family of transforming growth factor beta pathway components.
• Proc Natl Acad Sci U S A. 1996; 93: 790-4
• Display abstract

• Although transforming growth factor beta (TGF-beta) superfamily ligands play critical roles in diverse developmental processes, how cells transduce signals from these ligands is still poorly understood. Cell surface receptors for these ligands have been identified, but their cytoplasmic targets are unknown. We have identified three Caenorhabditis elegans genes, sma-2, sma-3, and sma-4, that have mutant phenotypes similar to those of the TGF-beta-like receptor gene daf-4, indicating that they are required for daf-4-mediated developmental processes. We show that sma-2 functions in the same cells as daf-4, consistent with a role in transducing signals from the receptor. These three genes define a protein family, the dwarfins, that includes the Mad gene product, which participates in the decapentaplegic TGF-beta-like pathway in Drosophila [Sekelsky, J. J., Newfeld, S. J., Raftery, L. A., Chartoff, E. H. & Gelbart, W. M. (1995) Genetics 139, 1347-1358]. The identification of homologous components of these pathways in distantly related organisms suggests that dwarfins may be universally required for TGF-beta-like signal transduction. In fact, we have isolated highly conserved dwarfins from vertebrates, indicating that these components are not idiosyncratic to invertebrates. These analyses suggest that dwarfins are conserved cytoplasmic signal transducers.

• ten Dijke P, Miyazono K, Heldin CH
• Signaling via hetero-oligomeric complexes of type I and type II serine/threonine kinase receptors.
• Curr Opin Cell Biol. 1996; 8: 139-45
• Display abstract

• Members of the transforming growth factor-beta (TGF-beta) superfamily have been found to signal by inducing the formation of hetero-oligomeric complexes of different type I and type II serine/threonine kinase receptors. Recent data indicate that binding of TGF-beta to its constitutively active type II receptor recruits the type I receptor into the complex; the type I receptor is thereafter phosphorylated and activated, processes which are necessary and sufficient for most TGF-beta mediated responses. Recent genetic analyses of Drosophila also indicate a strict requirement for both type I and type II receptors in decapentaplegic signaling in vivo.

• Hirai R, Fijita T
• A human transforming growth factor-beta type II receptor that contains an insertion in the extracellular domain.
• Exp Cell Res. 1996; 223: 135-41
• Display abstract

• In the signal transduction of transforming growth factor-beta (TGF-beta), the TGF-beta type II receptor (betaR-II) binds TGF-beta, presents the ligand to type I receptor, and forms a heterodimeric receptor complex that functions as an active signaling receptor. We isolated cDNA clones encoding an isoform of human betaR-II from vascular endothelial cells by RT-PCR. The deduced structure of the isoform designated as betaR-IIb contained a 25-amino-acid residue inserted at the extracellular region. The insert exhibited 73% homology to a similar insertion found in a mouse betaR-II variant. BetaR-IIb mRNA was ubiquitously expressed in all human cell lines examined. BetaR-IIb as well as betaR-II stably expressed in betaR-II deficient DR26 cells acted as a functional receptor transducing signals from TGF-beta for growth inhibition and the transcriptional activation of the plasminogen activator inhibitor-1 promoter. These results indicated that the betaR-IIb expressed in mammalian cells functions indistinguishably from in betaR-II. The potential function of betaR-IIb is discussed.

• Yingling JM, Das P, Savage C, Zhang M, Padgett RW, Wang XF
• Mammalian dwarfins are phosphorylated in response to transforming growth factor beta and are implicated in control of cell growth.
• Proc Natl Acad Sci U S A. 1996; 93: 8940-4
• Display abstract

• The dwarfin protein family has been genetically implicated in transforming growth factor beta (TGF-beta)-like signaling pathways in Drosophila and Caenorhabditis elegans. To investigate the role of these proteins in mammalian signaling pathways, we have isolated and studied two murine dwarfins, dwarfin-A and dwarfin-C. Using antibodies against dwarfin-A and dwarfin-C, we show that these two dwarfins and an immunogenically related protein, presumably also a dwarfin, are phosphorylated in a time- and dose-dependent manner in response to TGF-beta. Bone morphogenetic protein 2, a TGF-beta superfamily ligand, induces phosphorylation of only the related dwarfin protein. Thus, TGF-beta superfamily members may use overlapping yet distinct dwarfins to mediate their intracellular signals. Furthermore, transient overexpression of either dwarfin-A or dwarfin-C causes growth arrest, implicating the dwarfins in growth regulation. This work provides strong biochemical and preliminary functional evidence that dwarfin-A and dwarfin-C represent prototypic members of a family of mammalian proteins that may serve as mediators of signaling pathways for TGF-beta superfamily members.

• Quirin N, Feige JJ, Zaoui P
• [TGF-beta: usage in nephrology]
• Nephrologie. 1996; 17: 227-35
• Display abstract

• TGF beta is a ubiquitous multifunctional cytokine produced by a wide variety of cells; its amino acid sequence indicates a marked conservation across species. Five isoforms have been identified, three of which, TGF beta 1, 2, 3, in mammals. It is secreted in a biologically latent form and is then activated by physiological mechanisms which are still poorly characterized. TGF beta is known as a regulator of cell growth, differentiation and proliferation, but exerts also a tight control over extracellular matrix synthesis. Although TGF beta has been found to have a beneficial role in connective tissue and bone repair processes, its overexpression could lead to fibrosis and fibroproliferative diseases. TGF beta is also involved in kidney fibrogenesis and new pathways aimed at preventing glomerulosclerosis and interstitial fibrosis using TGF beta-inhibitors are new topics of research.

• Charng MJ, Kinnunen P, Hawker J, Brand T, Schneider MD
• FKBP-12 recognition is dispensable for signal generation by type I transforming growth factor-beta receptors.
• J Biol Chem. 1996; 271: 22941-4
• Display abstract

• The FK506-binding protein, FKBP12, is a putative target of type I receptors for transforming growth factor-beta (TbetaR-I). As the FK506 motif that competes with TbetaR-I for FKBP12 resembles an invariant Leu-Pro dipeptide in TbetaR-I, we replaced Leu193 and Pro194 with Ala, along with mutations across the Gly/Ser box. L193A, P194A, and L193A/P194A do not alter TbetaR-I function; T204D partially activates, independent of ligand; L193A/P194A/T204D was an even more potent constitutive mutation. Association with FKBP12 in a yeast two-hybrid assay was disrupted by P194A, L193A/P194A, and L193A/P194A/T204D, but not L193A or T204D alone. Thus, FKBP12 recognition is dispensable for TGFbeta signaling.

• Reilly KM, Melton DA
• Short-range signaling by candidate morphogens of the TGF beta family and evidence for a relay mechanism of induction.
• Cell. 1996; 86: 743-54
• Display abstract

• The specification and patterning of cell fates by a morphogen gradient is a unifying theme of developmental biology, yet little evidence exists for the presence of gradients in vivo or to show how such putative gradients form. Vg1 and activin are candidate morphogens involved in Xenopus mesoderm induction. This study suggests that these TGF beta family members act on adjacent cells but do not travel through the intact extracellular space to induce distant cells directly. Moreover, we present evidence for the presence of secondary inducing signals that could be involved in relaying signals to distant cells. These results suggest that if a localized cellular source of an inducer acts to pattern mesodermal cells at a distance in Xenopus embryos, it does so by a relay mechanism.

• Chen X, Rubock MJ, Whitman M
• A transcriptional partner for MAD proteins in TGF-beta signalling.
• Nature. 1996; 383: 691-6
• Display abstract

• The transforming-growth-factor-beta (TGF-beta) superfamily is critical for establishing mesoderm during early embryogenesis in Xenopus. The transcriptional activation of Mix.2, an immediate-early response gene specific to activin-like members of the TGF-beta superfamily, is associated with the rapid appearance of a site-specific DNA-binding activity that recognizes a fifty-base-pair regulatory element known as ARE in the Mix.2 promoter. Cloning of the site-specific DNA-binding component of this activity revealed it to be a new winged-helix transcription factor and a direct target for signalling by the TGF-beta superfamily. XMAD2, a recently identified TGF-beta signal transducer, forms a complex with the transcription factor in an activin-dependent fashion to generate an activated ARE-binding complex. A model is proposed to explain how TGF-beta superfamily signals might regulate the expression of specific genes in the early embryo.

• Mizumoto Y
• [Changes in NK activities and TGF- beta concentrations in the peritoneal cavity in endometriosis and their interaction related with infertility]
• Nippon Sanka Fujinka Gakkai Zasshi. 1996; 48: 379-85
• Display abstract

• The purpose of this study was to clarify the relationship between NK activity and TG-beta in the immune system in endometriosis. We investigated (1) the changes in the NK activity and concentration of TGF-beta in human peritoneal fluid (HPF), and (2) the effects of HPF and TGF-beta on the development of early mice embryos. In a rat model of experimental endometriosis, we observed the effects of tissue culture supernatants of peritoneum on NK activity in rat spleen cells, and obtained the following results. (1) NK activity of peripheral lymphocytes in healthy women was significantly suppressed in the presence of HPF of endometriosis. (2) The concentrations of TGF-beta was significantly higher in HPF of endometriosis than in HPF of healthy women. (3) Both HPF of endometriosis and TGF-beta significantly inhibited the development of early mice embryos. (4) The supernatants prepared from the intact peritoneum of the rat model showed marked inhibition of NK activity compared to control rats, although the peritoneum was obtained from a region distant from the implanted endometrium. These results suggest that ectopic endometrial tissues may cause a change in the cell-mediated immune system and subsequently exert an adverse effect on human reproduction.

• Zhang Y, Feng X, We R, Derynck R
• Receptor-associated Mad homologues synergize as effectors of the TGF-beta response.
• Nature. 1996; 383: 168-72
• Display abstract

• Transforming growth factor-beta TGF-beta is the prototype for a family of extracellular proteins that affect cell proliferation and tissue differentiation. TGF-beta-related factors, including BMP-2/4, Dpp and activin, act through two types of serine/threonine kinase receptors which can form a heteromeric complex. However, the mechanism of signal transduction by these receptors is largely unknown. In Drosophila, Mad is required for signalling by Dpp. We have isolated complementary DNAs for four human Mad homologues, one of which, hMAD-4, is identical to DPC-4, a candidate tumour suppressor. hMAD-3 and -4 synergized to induce strong ligand-independent TGF-beta-like responses. When truncated at their carboxy termini, hMAD-3 and -4 act as dominant-negative inhibitors of the normal TGF-beta response. The activity of hMAD-3 and -4 was regulated by the TGF-beta receptors, and hMAD-3 but not hMAD-4 was phosphorylated and associated with the ligand-bound receptor complex. These results define hMAD-3 and -4 as effectors of the TGF-beta response and demonstrate a function for DPCA-4/hMAD-4 as a tumour suppressor.

• Diaz-Ruiz C, Montaner B, Perez-Tomas R
• Immunochemical study of transforming growth factor-beta in the kidney of the rat and chicken.
• Histochem Cell Biol. 1996; 105: 475-8
• Display abstract

• Transforming growth factor-beta (TGF-beta) is a homodimeric polypeptide of 25 kDa, which regulates cell growth and differentiation and influences extracellular matrix metabolism. Using immunochemical techniques, we identified TGF-beta in the loops of Henle and the collecting and Bellini ducts of rat kidney and in the loops of Henle of chicken kidney. Furthermore, we detected two TGF-beta-immunoreactive proteins on kidney blots of the rat of 12.5 and 47 kDa, and three on chicken kidney blots of 12.5, 34, and 47 kDa. We suggest that the precursor forms of rat and chicken TGF-beta 2 or beta 3, chicken TGF-beta 4, and the mature form of all of them are expressed in the collecting and Bellini ducts of rat kidney and the loops of Henle of rat and chicken kidney.

• Francis-West P, Tickle C
• Limb development.
• Curr Top Microbiol Immunol. 1996; 212: 239-59

• Riggins GJ et al.
• Mad-related genes in the human.
• Nat Genet. 1996; 13: 347-9
• Display abstract

• Resistance to the growth inhibitory effects of TGF-beta is common in human cancers. However, the mechanism(s) by which tumour cells become resistant to TGF-beta are generally unknown. We have identified five novel human genes related to a Drosophila gene called Mad which is thought to transduce signals from TGF-beta family members. One of these genes was found to be somatically mutated in two of eighteen colorectal cancers, and three of the other genes were located at chromosomal positions previously suspected to harbor tumour suppressor genes. These data suggest that this gene family may prove to be important in the suppression of neoplasia, imparting the growth inhibitory effects of TGF-beta-like ligands.

• Derynck R et al.
• Nomenclature: vertebrate mediators of TGFbeta family signals.
• Cell. 1996; 87: 173-173

• Kaneko H, Horiike S, Taniwaki M, Misawa S
• Microsatellite instability is an early genetic event in myelodysplastic syndrome but is infrequent and not associated with TGF-beta receptor type II gene mutation.
• Leukemia. 1996; 10: 1696-9
• Display abstract

• We examined microsatellite instability (MSI) at 10 loci of dinucleotide repeats using the polymerase chain reaction (PCR) in patients with myelodysplastic syndrome (MDS). Bone marrow DNA was obtained from 45 patients repeatedly during the disease course and fibroblast DNA was also collected from 19 of them as a normal control. Three of the 19 patients showed an alteration at more than three loci, when the allele length was compared between their fibroblast DNA and the initial marrow DNA. On the other hand, none of the 45 patients showed an alteration when the initial sample was compared with the latest one. One of the three patients with MSI had refractory anemia and two refractory anemia with ring sideroblasts and none of them showed disease progression, complex chromosome abnormality, karyotypic evolution, or mutation of N-RAS or TP53. Moreover, a frameshift mutation within 10 repeating adenines of transforming growth factor beta type II receptor gene, which was recently recognized as a critical target of MSI, was not found in any of the patients including the three with MSI. These findings suggest that MSI is an early but infrequent genetic event and is independent of other critical genetic aberrations in the development of MDS.

• Levy LS, Bost KL
• Mechanisms that contribute to the development of lymphoid malignancies: roles for genetic alterations and cytokine production.
• Crit Rev Immunol. 1996; 16: 31-57
• Display abstract

• Recent studies have defined genetic alterations commonly associated with transformed lymphocytes. This review suggests roles for these alterations in the development of lymphoid neoplasms. Damage to the genes encoding proteins that function in intracellular signaling, transcription, or regulation of the cell cycle has been identified and linked at varying degrees to the progression of certain lymphoid malignancies. An understanding of the mechanistic consequences following such genetic alterations is essential to an understanding of the development of these lymphoid neoplasms. In contrast, it is also becoming clear that the dysregulated expression of proteins that are not genetically altered can also contribute to the progression of lymphoid malignancies. One such example is the excessive expression of "normal" lymphokines of cytokines which accompanies many lymphoproliferative diseases. The dysregulated expression of cytokines during malignancy can result in the augmentation of growth of transformed lymphocytes, as well as an alteration of the anti-tumor immune response. The latter mechanism is especially important because evasion of the impending immune response is a prerequisite for the progression of lymphoproliferative diseases. Taken together, this review supports the notion that the development of lymphoid malignancies is multifactorial, involving genetic alterations as well as dysregulated cytokine expression.

• Wright JA, Huang A
• Growth factors in mechanisms of malignancy: roles for TGF-beta and FGF.
• Histol Histopathol. 1996; 11: 521-36
• Display abstract

• Malignant progression is a complex process involving the accumulation of multiple genetic alterations leading to changes in many specialized cell functions. Important in this process is the loss of growth control which is frequently associated with modifications in growth factor production, and growth factor response pathways. Indeed, oncogenes have been characterized that code for polypeptide growth factors or their receptors, and many tumor cell populations release potently mitogenic growth factors which contribute to the malignant properties of tumor cells. In this review, the importance of growth factors in mechanisms of malignant progression is emphasized, using as examples the transforming growth factor-beta (TGF-beta) and fibroblast growth factor (FGF) families. We describe many of the properties and biological activities of these two families of growth factors, focusing on mechanisms of autocrine and intracrine mitogenic stimulation of tumor cell proliferation and malignant progression. The discussion includes evidence for altered growth factor expression in tumor cells, and the relationship between these changes in growth factors and alterations in the regulation of DNA synthesis, cell proliferation, protease production and cell motility required for invasion and metastasis. Recent studies are described that show that aberrant expression of TGF-beta1, bFGF or K-FGF results in dramatic changes in the genetic stability of cells, leading to increased rates of spontaneous gene amplification and the generation of drug resistant variants. These findings describe new malignancy relevant functions for altered growth factor expression.

• Glinka A, Delius H, Blumenstock C, Niehrs C
• Combinatorial signalling by Xwnt-11 and Xnr3 in the organizer epithelium.
• Mech Dev. 1996; 60: 221-31
• Display abstract

• The epithelium of the Spemann organizer plays an important role in embryonic axis formation and transplantation experiments have shown that epithelial organizer cells have potent axis-inducing potential. Known axis-inducing molecules like noggin and chordin are not expressed in the epithelium and cannot account for its inductive properties. Xwnt-11 is expressed in the epithelium but has only poor dorsalizing activity. In an expression screen for genes that are able to functionally cooperate with Xwnt-11 we have identified a cDNA encoding Xenopus nodal-related 3 (XNR3), a member of the TGF-beta family, coexpressed with Xwnt-11 in the organizer epithelium. Xwnt-11 and Xnr3 act highly cooperatively in inducing secondary embryonic axes and dorsalizing ventral mesoderm. Xwnt-11/Xnr3 interfere with BMP signalling without themselves inducing chordin or noggin. The results indicate that induction by the organizer epithelium may result from the combinatorial action of instructive Xnr3 and permissive Xwnt-11 signalling.

• Eppert K et al.
• MADR2 maps to 18q21 and encodes a TGFbeta-regulated MAD-related protein that is functionally mutated in colorectal carcinoma.
• Cell. 1996; 86: 543-52
• Display abstract

• The MAD-related (MADR) family of proteins are essential components in the signaling pathways of serine/threonine kinase receptors for the transforming growth factor beta (TGFbeta) superfamily. We demonstrate that MADR2 is specifically regulated by TGFbeta and not bone morphogenetic proteins. The gene for MADR2 was found to reside on chromosome 18q21, near DPC4, another MADR protein implicated in pancreatic cancer. Mutational analysis of MADR2 in sporadic tumors identified four missense mutations in colorectal carcinomas, two of which display a loss of heterozygosity. Biochemical and functional analysis of three of these demonstrates that the mutations are inactivating. These findings suggest that MADR2 is a tumor suppressor and that mutations acquired in colorectal carcinomas may function to disrupt TGFbeta signaling.

• Duh QY, Grossman RF
• Thyroid growth factors, signal transduction pathways, and oncogenes.
• Surg Clin North Am. 1995; 75: 421-37
• Display abstract

• Growth of thyroid cancer cells is stimulated by various growth factors via signal transduction pathways. TSH, EGF, IGF, and TGF-alpha stimulate and TGF-beta inhibits thyroid cell growth. TSH stimulates thyroid cells via both the adenylate cyclase-PKA and the PLC-PKC-Ca signal transduction pathways. TSH-r, ras, gsp, ret, trk, and myc are oncogenes that are activated in some thyroid neoplasms. P53 and RB are tumor suppressor genes that are inactivated in some thyroid cancers.

• Marx J
• DNA repair defect tied to mutated TGF-beta receptor gene.
• Science. 1995; 268: 1276-7

• Kumano K, Shimoda M, Hyodo T, Sakai T
• The role of TGF-beta in growth inhibition of peritoneal mesothelial cells in high-glucose dialysate.
• Perit Dial Int. 1995; 15: 935-935

• Sosroseno W, Herminajeng E
• The immunoregulatory roles of transforming growth factor beta.
• Br J Biomed Sci. 1995; 52: 142-8
• Display abstract

• Transforming growth factor-beta is a 25 kD polypeptide produced by both healthy and neoplastic cells. This cytokine effects the introduction of immune response, but its exact mechanism remains to be elucidated. In this paper the immuno-regulatory role of this cytokine on both the immunocompetent cells and accessory molecules is discussed. The growth factor-induced up and down regulation of the immune response suggest that transforming growth factor-beta can done some extent modulate certain pathological conditions, and hence suppressed production of this growth factor has potential benefits for bimodal immunotherapy.

• Schneider MD, Brand T
• Molecular analysis of TGF beta signal transduction. Dominant-inhibitory mutations of the type II and type I TGF beta receptor.
• Ann N Y Acad Sci. 1995; 752: 309-16

• Wang YQ, Sizeland A, Wang XF, Sassoon D
• Restricted expression of type-II TGF beta receptor in murine embryonic development suggests a central role in tissue modeling and CNS patterning.
• Mech Dev. 1995; 52: 275-89
• Display abstract

• The type-II TGF beta receptor mediates many of the biological responses to TGF beta. An examination of the expression of the type-II TGF beta receptor during mouse embryogenesis therefore provides specific information about the role of TGF beta during embryogenesis than has been available to date. We have isolated the genomic murine homologue of the human type-II TGF beta receptor corresponding to exon 2. The murine and human sequences show a high degree of homology. Using the murine probe, we found that type-II TGF beta receptor expression is regulated in both a spatial and a temporal fashion by using in situ hybridization and ribonuclease protection assays. Type-II TGF beta receptor expression is localized to the mesenchyme during critical interactions with adjacent epithelium such as developing hair follicles, whisker follicles and tooth anlage. In the central nervous system, type-II TGF beta receptor expression is highly restricted to the floor plate. Strong expression is also detected in migrating neural crest cells, meninges, and choroid plexus. Specific mesenchymal localization of type-II TGF beta receptor is also observed in lung, kidney, intestine, stomach, and bladder. The restricted expression of type-II TGF beta receptor in mesenchymal cells at sites of epithelial-mesenchymal interactions suggests that type-II TGF beta receptor plays a major role in mediating the establishment of embryonic organ systems. The highly restricted expression of type-II TGF beta receptor in the developing CNS suggests an important role for a serine/threonine kinase in patterning of the nervous system.

• Yamaguchi A
• Regulation of differentiation pathway of skeletal mesenchymal cells in cell lines by transforming growth factor-beta superfamily.
• Semin Cell Biol. 1995; 6: 165-73
• Display abstract

• The skeletal mesenchymal cells including osteoblasts, chondrocytes, muscle cells and adipocytes are believed to be derived from the common mesenchymal progenitors. Using various cell lines, multipotential mesenchymal progenitors capable of differentiating into these cell lineages have been demonstrated to be present in the skeletal tissues. The developmental potential of the progenitors becomes more restricted according to their maturation stages in the specialized cell lineages. Among the various growth factors, bone morphogenetic protein (BMP), which belongs to the transforming growth factor-beta (TGF-beta) superfamily, plays an important role in the regulation of the differentiation pathway of these mesenchymal cells. BMP-2 induces the undifferentiated mesenchymal progenitors to differentiate into osteoblasts, chondrocytes and adipocytes. BMP-2 also stimulates the committed osteo-chondroprogenitors to differentiate into more mature cell types, but inhibits myogenic cells to differentiate into myotubes. BMP-2 converts the differentiation pathway of C2C12 myoblasts into the osteogenic lineage associating with decreases in the expression levels of the muscle specific regulatory factors such as myogenin and MyoD. TGF-beta inhibits myotube formation in myogenic cell lines, but does not stimulate osteoblast differentiation. The TGF-beta superfamily plays important roles in the differentiation process of the skeletal mesenchymal cells.

• Ecochard V, Cayrol C, Foulquier F, Zaraisky A, Duprat AM
• A novel TGF-beta-like gene, fugacin, specifically expressed in the Spemann organizer of Xenopus.
• Dev Biol. 1995; 172: 699-703
• Display abstract

• Using a differential screening strategy, we have cloned a novel Xenopus gene, fugacin, related to the transforming growth factor beta superfamily. Transcripts were detected primarily in the dorsal marginal zone of late blastula. Thereafter, they became highly localized to the blastopore lip of early gastrula and were not observed at later stages. This gene, which is most homologous to the mouse gene nodal, displays a new pattern of cysteine residues. These findings highlight the potential role of these growth factors during early vertebrate development.

• Newfeld SJ, Gelbart WM
• Identification of two Drosophila TGF-beta family members in the grasshopper Schistocerca americana.
• J Mol Evol. 1995; 41: 155-60
• Display abstract

• Intercellular signaling molecules of the transforming growth factor-beta (TGF-beta) superfamily are required for pattern formation in many multicellular organisms. The decapentaplegic (dpp) gene of Drosophila melanogaster has several developmental roles. To improve our understanding of the evolutionary diversification of this large family we identified dpp in the grasshopper Schistocerca americana. S. americana diverged from D. melanogaster approximately 350 million years ago, utilizes a distinct developmental program, and has a 60-fold-larger genome than D. melanogaster. Our analyses indicate a single dpp locus in D. melanogaster and S. americana, suggesting that dpp copy number does not correlate with increasing genome size. Another TGF-beta superfamily member, the D. melanogaster gene 60A, is also present in only one copy in each species. Comparison of homologous sequences from D. melanogaster, S. americana, and H. sapiens, representing roughly 900 million years of evolutionary distance, reveals significant constraint on sequence divergence for both dpp and 60A. In the signaling portion of the dpp protein, the amino acid identity between these species exceeds 74%. Our results for the TGF-beta superfamily are consistent with current hypotheses describing gene duplication and diversification as a frequent response to high levels of selective pressure on individual family members.

• Stavnezer J
• Regulation of antibody production and class switching by TGF-beta.
• J Immunol. 1995; 155: 1647-51

• Yamaguchi K et al.
• Identification of a member of the MAPKKK family as a potential mediator of TGF-beta signal transduction.
• Science. 1995; 270: 2008-11
• Display abstract

• The mitogen-activated protein kinase (MAPK) pathway is a conserved eukaryotic signaling module that converts receptor signals into various outputs. MAPK is activated through phosphorylation by MAPK kinase (MAPKK), which is first activated by MAPKK kinase (MAPKKK). A genetic selection based on a MAPK pathway in yeast was used to identify a mouse protein kinase (TAK1) distinct from other members of the MAPKKK family. TAK1 was shown to participate in regulation of transcription by transforming growth factor-beta (TGF-beta). Furthermore, kinase activity of TAK1 was stimulated in response to TGF-beta and bone morphogenetic protein. These results suggest that TAK1 functions as a mediator in the signaling pathway of TGF-beta superfamily members.

• Raftery LA, Twombly V, Wharton K, Gelbart WM
• Genetic screens to identify elements of the decapentaplegic signaling pathway in Drosophila.
• Genetics. 1995; 139: 241-54
• Display abstract

• Pathways for regulation of signaling by transforming growth factor-beta family members are poorly understood at present. The best genetically characterized member of this family is encoded by the Drosophila gene decapentaplegic (dpp), which is required for multiple events during fly development. We describe here the results of screens for genes required to maximize dpp signaling during embryonic dorsal-ventral patterning. Screens for genetic interactions in the zygote have identified an allele of tolloid, as well as two novel alleles of screw, a gene recently shown to encode another bone morphogenetic protein-like polypeptide. Both genes are required for patterning the dorsalmost tissues of the embryo. Screens for dpp interactions with maternally expressed genes have identified loss of function mutations in Mothers against dpp and Medea. These mutations are homozygous pupal lethal, engendering gut defects and severely reduced imaginal disks, reminiscent of dpp mutant phenotypes arising during other dpp-dependent developmental events. Genetic interaction phenotypes are consistent with reduction of dpp activity in the early embryo and in the imaginal disks. We propose that the novel screw mutations identified here titrate out some component(s) of the dpp signaling pathway. We propose that Mad and Medea encode rate-limiting components integral to dpp pathways throughout development.

• Massague J, Polyak K
• Mammalian antiproliferative signals and their targets.
• Curr Opin Genet Dev. 1995; 5: 91-6
• Display abstract

• The effect of mitogens on the mammalian cell cycle is opposed by the action of antimitogens, such as TGF-beta, cAMP agonists, and various antiproliferative drugs. The recent identification of TGF-beta receptors that initiate antimitogenic signals and of cell cycle kinase inhibitors that respond to these signals has provided new insights into this process. The evidence argues that mitogenic and antimitogenic signals confront each other by regulating in opposite ways the activity of cyclin-dependent kinases that control cell commitment to DNA replication.

• Hall BK, Miyake T
• Divide, accumulate, differentiate: cell condensation in skeletal development revisited.
• Int J Dev Biol. 1995; 39: 881-93
• Display abstract

• Cell condensation is a pivotal stage in skeletal development. Although prechondrogenic condensations normally exist for some 12 h, duration can vary. Variation is seen both between condensations for different cartilages (Meckel's vs. elastic ear cartilage) and within a single condensation from which more than one skeletal element will form, as in the three components of the single first arch chondrogenic condensation. Understanding how duration of the condensation phase is established--how the condensation phase is entered and exited during cell differentiation--remains a major area for future study. During chondrogenesis, cell-specific products such as collagen types II and IX and cartilage proteoglycan appear concomitant with condensation. Therefore, during chondrogenesis, condensation precedes commitment of cells as prechondroblasts. During osteogenesis, however, differentiation of preosteoblasts precedes condensation. Therefore, during osteogenesis, condensation amplifies the number of committed osteogenic cells. Further comparative analysis of skeletogenesis should provide us with a more rigorous understanding of cell commitment, when differentiation is initiated, how commitment and differentiation are measured and the relationship of condensation to onset of differentiation. Current knowledge of molecules characteristic of condensations focused attention on extracellular matrix and cell surface components on the one hand, and on growth factors homeobox genes and transcription factors on the other. We have drawn together the molecular data for pre-chondrogenic condensations in diagrammatic form in Figure 2. Three major phases of chondrogenesis are identified: (a) epithelial-mesenchymal interactions that precede condensation, (b) condensation itself, and (c) cell differentiation. Although we label the third phase differentiation, it is important to recognize that phases a and b also constitute aspects of chondroblast cell differentiation (see Dunlop and Hall, 1995 for a discussion of this point. The pre-condensation phase is characterized by expression of Hox genes, growth factors (TGF-beta and BMP-2) and the cell surface proteoglycan receptor, syndecan-1. Expression of Msx-1 and Msx-2, growth factors and syndecan continues into the condensation phase. Other molecules, such as versican, syndecan-3 and tenascin, present in low concentrations before condensation, are up-regulated during condensation. Yet other molecules--Hox genes, transcription factors, growth factors (activin, BMP-4 and -5, GDF-5), cell adhesion molecules and proteoglycans--are only expressed during the condensation phase, while the transcription factor Pax-1, fibronectin, hyaluronan and hyaladherin are expressed both during and after condensation. During condensation mRNAs for collagen types II and IX and for the core protein of cartilage proteoglycan are up-regulated. Late in condensation and increasingly thereafter, the protein products of these genes accumulate as chondroblasts differentiate (see Fig. 2 for details). Not all the molecules present before, during of after condensation can be placed into causal sequences. Some however can. In Figure 3 we summarize the causal sequences discussed in this paper as they relate to initiation of condensation and to transit from condensation to overt differentiation during chondrogenesis. Condensations form following activation of at least three pathways: (1) Initiation of epithelial-mesenchymal interactions by tenascin, BMP-2, TGF beta-1 and Msx-1 and -2. (2) Up-regulation of N-CAM by activin. (3) Up-regulation of fibronectin by TGF-beta, further enhancing N-CAM accumulation (Fig. 3). It is by these three pathways that condensations are initiated and grow. Transition from condensation to overt cell differentiation is under both positive and negative control (Fig. 3). Syndecan blocks fibronectin and so blocks N-CAM accumulation, preventing accumulation of additional cell

• Sun D, Kar S, Carr BI
• Differentially expressed genes in TGF-beta 1 sensitive and resistant human hepatoma cells.
• Cancer Lett. 1995; 89: 73-9
• Display abstract

• The pathway of growth inhibitory cellular signal transduction by transforming growth factor beta 1 is largely unknown. Although several cellular proteins have been shown to be involved in the pathway, others remain to be identified. To search for other involved proteins, differentially expressed genes were examined in two human hepatoma cell lines that were respectively sensitive and resistant to growth inhibition by TGF-beta 1. Two such genes (SB31 and SB16) were characterized and found to be 100% homologous to leucine-rich alpha 2-glycoprotein and the ribosomal protein S25, respectively. SB31 was coordinately expressed with the TGF-beta I type II receptor, implicating a possible interaction. Expression of both genes in different cell lines can be broadly correlated with the sensitivity of the cell lines to growth inhibition by TGF-beta 1. SB16 expression is strongly suppressed in rat liver regeneration after partial hepatectomy, suggesting that it may have a role in liver growth.

• Johnston JB et al.
• Deposition of transforming growth factor-beta in the marrow in myelofibrosis, and the intracellular localization and secretion of TGF-beta by leukemic cells.
• Am J Clin Pathol. 1995; 103: 574-82
• Display abstract

• The marrows of 10 patients with hematologic malignancies were examined by immunohistochemistry using anti TGF-beta antibody, CC(1-30), which detects secreted TGF-beta, and compared with four normal marrows. TGF-beta was not demonstrated in marrows with a normal level of reticulin fibrosis; however, TGF-beta was observed within collagen in marrows having collagen fibrosis or increased reticulin fibrosis. The extent of TGF-beta deposition paralleled the severity of fibrosis (P < .0001), and occurred even with normal or reduced numbers of megakaryocytes. Using another TGF-beta antibody, LC(1-30), which detects intracellular TGF-beta, TGF-beta was detected by immunofluorescence in discrete sites in the cytoplasm of immature and mature myeloid and large granular lymphocytic leukemia cells. These sites colocalized with areas detected by an anti-granule antibody (D545) suggesting that TGF-beta was stored in granules. However, neither the TGF-beta mRNA content nor the degree of TGF-beta secretion by these leukemic cells correlated with the extent of TGF-beta deposition in the marrow. Thus, TGF-beta deposition in marrow may contribute to myelofibrosis, but the source of this cytokine in the absence of megakaryocytes requires further study.

• Smith RA
• The effect on TGF-beta 1 on osseointegration.
• J Calif Dent Assoc. 1995; 23: 49-53
• Display abstract

• Conventional techniques for placement of endosseous implants may take more than one year for extraction, healing, implant placement and restoration. Although clinical success rates are reported to be greater than 90 percent, failures do occur, in part because of incomplete osseointegration at the implant-bone interface. We hypothesized that implants placed in the standard fashion into extraction sockets and edentulous ridges of mature minipigs treated with human recombinant Transforming Growth Factor (TGF-beta 1), a multifunctional regulatory protein, will achieve a more rapid and higher percentage surface of osseointegration than in comparable sites without TGF-beta 1. Our results are consistent with the hypothesis that TGF-beta 1 is an osteoinductive factor in vivo enhancing the quantity of osseointegration of endosseous implants. However, the rate of osseointegration was not accelerated.

• Brand T, Schneider MD
• The TGF beta superfamily in myocardium: ligands, receptors, transduction, and function.
• J Mol Cell Cardiol. 1995; 27: 5-18

• Metcalfe JC, Grainger DJ
• TGF-beta: implications for human vascular disease.
• J Hum Hypertens. 1995; 9: 679-679

• Wharton KA
• How many receptors does it take?
• Bioessays. 1995; 17: 13-6
• Display abstract

• Three recent reports identify two genes, thick veins (tkv) and saxophone (sax), which encode serine/threonine transmembrane proteins that act as receptors for mediating different aspects of the Drosophila TGF-beta-related signal, dpp. tkv is required for patterning the entire embryonic dorsal region, while sax is required for patterning only amnioserosa, the dorsalmost cell fates. dpp signaling in other developmental processes again requires both tkv and sax, but to differing degrees. tkv and sax encode type I receptors, which appear to directly bind ligand, unlike what has been observed for other type I receptors.

• Letsou A et al.
• Drosophila Dpp signaling is mediated by the punt gene product: a dual ligand-binding type II receptor of the TGF beta receptor family.
• Cell. 1995; 80: 899-908
• Display abstract

• Signaling by TGF beta-related factors requires ligand-induced association between type I and type II transmembrane serine/threonine kinases. In Drosophila, the saxophone (sax) and thick veins (tkv) genes encode type I receptors that mediate signaling by decapentaplegic (dpp), a member of the bone morphogenetic protein (BMP) subgroup of TGF beta-type factors. In this report, we demonstrate that the Drosophila punt gene encodes atr-II, a previously described type II receptor that on its own is able to bind activin but not BMP2, a vertebrate ortholog of dpp. Mutations in punt produce phenotypes similar to those exhibited by tkv, sax, and dpp mutants. Furthermore, punt will bind BMP2 in concert with tkv or sax, forming complexes with these receptors. We suggest that punt functions as a type II receptors for dpp and propose that BMP signaling in vertebrates may also involve sharing of type II receptors by diverse ligands.

• Staehling-Hampton K, Laughon AS, Hoffmann FM
• A Drosophila protein related to the human zinc finger transcription factor PRDII/MBPI/HIV-EP1 is required for dpp signaling.
• Development. 1995; 121: 3393-403
• Display abstract

• Little is known about the signal transduction pathways by which cells respond to mammalian TGF-beta s or to decapentaplegic (dpp), a Drosophila TGF-beta-related factor. Here we describe the genetic and molecular characterization of Drosophila schnurri (shn), a putative transcription factor implicated in dpp signaling. The shn protein has eight zinc fingers and is related to a human transcription factor, PRDII/MBPI/HIV-EP1, that binds to nuclear factor-kappa B-binding sites and activates transcription from the HIV long terminal repeat (LTR). shn mRNA is expressed in a dynamic pattern in the embryo that includes most of the known target tissues of dpp, including the dorsal blastoderm, the mesodermal germlayer and parasegments 4 and 7 of the midgut. Mutations in shn affect several developmental processes regulated by dpp including induction of visceral mesoderm cell fate, dorsal/ventral patterning of the lateral ectoderm and wing vein formation. Absence of shn function blocks the expanded expression of the homeodomain protein bagpipe in the embryonic mesoderm caused by ectopic dpp expression, illustrating a requirement for shn function downstream of dpp action. We conclude that shn function is critical for cells to respond properly to dpp and propose that shn protein is the first identified downstream component of the signal transduction pathway used by dpp and its receptors.

• Pankratz MJ, Hoch M
• Control of epithelial morphogenesis by cell signaling and integrin molecules in the Drosophila foregut.
• Development. 1995; 121: 1885-98
• Display abstract

• Coordinated cell movements are critical for tissue and organ morphogenesis in animal development. We show that the Drosophila genes hedgehog and wingless, which encode signaling molecules, and the gene myospheroid, which encodes a beta subunit of the integrins, are required for epithelial morphogenesis during proventriculus development. In contrast, this morphogenetic process is suppressed by the decapentaplegic gene, which encodes a member of the TGF beta family of growth factors. These results identify a novel cell signaling center in the foregut that directs the formation of a multiply folded organ from a simple epithelial tube.

• Tiedemann H, Tiedemann H, Grunz H, Knochel W
• Molecular mechanisms of tissue determination and pattern formation in amphibian embryos.
• Naturwissenschaften. 1995; 82: 123-34
• Display abstract

• Factors of the TGF-beta superfamily (activin, vegetalizing factor) and the FGF family determine endoderm and mesoderm. The dorsoventral polarity of the mesoderm depends on additional factors (BMP-4, Wnt-8, noggin). Activin can directly activate gene transcription by signal transduction. Mesoderm is determined by factors prelocalized in the marginal zone. Its differentiation depends also on the animal ectoderm. Neural inducing factors have been partially purified. A masked neuralizing factor in the ectoderm is activated by induction of the ectoderm to the nervous system. Phorbolester can evoke neuralization signaling.

• Huang HC, Murtaugh LC, Vize PD, Whitman M
• Identification of a potential regulator of early transcriptional responses to mesoderm inducers in the frog embryo.
• EMBO J. 1995; 14: 5965-73
• Display abstract

• The activin/transforming growth factor-beta (TGF-beta) family of peptide growth factors plays a central role in the induction of mesoderm during early Xenopus embryogenesis. Immediate transcriptional responses to mesoderm-inducing signals have been described, but the signal transduction steps leading to these early responses are unknown. We describe here the first pre-transcriptional response to activin/TGF-beta mesoderm inducers in the early embryo. We have identified a cellular factor which binds to a 50 bp portion of the promoter for an activin/TGF-beta early response gene. This factor is activated within 4 min of treatment of embryonic prospective ectoderm with mesoderm-inducing factors, making it the earliest response to these factors described in early embryos. This factor can be activated throughout early cleavage and blastula stages, is activated by mesoderm inducers of the activin/TGF-beta superfamily but not the fibroblast growth factor (FGF) family, and does not appear to require an endogenous FGF signal for activation. Characterization of this factor provides a powerful tool for studying the early steps in the induction of mesoderm by members of the activin/TGF-beta superfamily.

• Rowe PM
• Clinical potential for TGF-beta.
• Lancet. 1994; 344: 72-3

• Miyazono K, Ten Dijke P, Ichijo H, Heldin CH
• Receptors for transforming growth factor-beta.
• Adv Immunol. 1994; 55: 181-220

• Chambaz EM, Cochet C, Souchelnitskiy S, Defaye G, Feige JJ
• Growth factors and pericellular components as a local signalling relay in the action of systemic hormones.
• Horm Res. 1994; 42: 35-8
• Display abstract

• Recent years have shown that peptidic signals, originally identified as growth factors, may represent a local signalling system involved in the regulation of cell function under the dependence of systemic hormones. As an example, (i) adrenocortical cells are highly sensitive to transforming growth factor-beta (TGF beta) which is a powerful inhibitor of their differentiated functions; (ii) adrenocortical cells produce and secrete TGF beta, which is present in the adrenal cortex in situ; (iii) adrenocorticotropin (ACTH), which is the best known positive effector of adrenocortical functions induces an up-regulation of TGF beta receptors in adrenocortical cells. Altogether, these observations suggest that TGF beta may be a local signalling system acting in balance with the systemic hormone ACTH to eventually tune the ability of adrenocortical cells to produce corticosteroids.

• Kim SJ, Romeo D, Yoo YD, Park K
• Transforming growth factor-beta: expression in normal and pathological conditions.
• Horm Res. 1994; 42: 5-8
• Display abstract

• Expression of the various isoforms of transforming growth factor-beta (TGF-beta) is differentially controlled both in vivo and in vitro. Characterization of the molecular mechanisms governing expression of TGF-beta isoforms now provides a basis for understanding the selective regulation of expression of the TGF-beta s by a variety of factors including oncogenes and tumor suppressor genes. In addition to transcriptional control, data suggest that expression of TGF-beta s is also regulated posttranscriptionally. Regulation of TGF-beta s by steroids and retinoids appears to involve predominantly posttranscriptional mechanisms. Identification of these mechanisms may contribute to the understanding of the regulatory events controlling cellular proliferation and differentiation by TGF-beta s and steroids/retinoids.

• Nugent P, Greene RM
• Interactions between the transforming growth factor beta (TGF beta) and retinoic acid signal transduction pathways in murine embryonic palatal cells.
• Differentiation. 1994; 58: 149-55
• Display abstract

• Regulation of expression of transforming growth factor-beta 3 (TGF-beta 3) and the cellular retinoic acid-binding proteins-I and II (CRABP-I, -II) by retinoic acid (RA) and TGF-beta was examined in primary cultures of murine embryonic palate mesenchymal (MEPM) cells. Northern blot hybridization revealed that RA and TGF-beta 1, beta 2 and beta 3 stimulated the expression of TGF-beta 3 mRNA within 24 hours of treatment. RA down-regulated the expression of CRABP-I mRNA and up-regulated the expression of CRABP-II mRNA in a time- and dose-dependent fashion. TGF-beta 1, beta 2 and beta 3 also down-regulated the expression of CRABP-I mRNA, while epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha) were without effect. TGF-beta 1 also stimulated a dose-dependent increase in the expression of CRABP-II mRNA. Again EGF and TGF-alpha were without effect. Basic fibroblast growth factor (bFGF) elicited a slight inhibitory effect on CRABP-II and a slight stimulatory effect on CRABP-I mRNA expression. Thus, cells derived from the mammalian developing palate express CRABP-I and CRABP-II mRNAs, both of which may be regulated by RA and TGF-beta. These data constitute the first demonstration of an effect of TGF-beta on the expression of CRABP-I and CRABP-II and provide further evidence for cross-talk between RA and TGF-beta signal transduction pathways.

• Schluesener HJ, Meyermann R
• Expression of BMP-6, a TGF-beta related morphogenetic cytokine, in rat radial glial cells.
• Glia. 1994; 12: 161-4
• Display abstract

• Bone morphogenetic protein-6 (BMP-6) is a member of the TGF-beta superfamily of cytokines. The bone morphogenetic proteins and homologous cytokines participate in realization of the genetic body plan by regulating homeotic gene expression, embryonic development, and neurogenesis. Here we demonstrate expression of BMP-6 in rat radial glial cells which are involved in embryonic organisation of the central nervous system. Thus, morphogenetic cytokines like BMP-6 might contribute to radial glial cell function in organizing the migration of immature neurons during the development of the CNS cortex.

• Okadome T, Yamashita H, Franzen P, Moren A, Heldin CH, Miyazono K
• Distinct roles of the intracellular domains of transforming growth factor-beta type I and type II receptors in signal transduction.
• J Biol Chem. 1994; 269: 30753-6
• Display abstract

• Transforming growth factor-beta (TGF-beta) transduces signals through binding to type I (T beta R-I) and type II (T beta R-II) serine/threonine kinase receptors. T beta R-I requires T beta R-II for ligand binding, whereas T beta R-II requires T beta R-I for signaling. We generated two different chimeric TGF-beta receptors, i.e. T beta R-1.2 containing the extracellular domain of T beta R-I and the intracellular domain of T beta R-II, and T beta R-2.1 containing the extracellular domain of T beta R-II and the intracellular domain of T beta R-I. T beta R-2.1 bound 125I-TGF-beta 1 alone, whereas T beta R-1.2 bound the ligand only in the presence of T beta R-II or T beta R-2.1. When transfected into a mutant mink lung epithelial cell line that lacks functional T beta R-II, T beta R-II cDNA, but not T beta R-2.1 cDNA, restored the responsiveness to TGF-beta 1 with regard to transcriptional activation of plasminogen activator inhibitor-1 gene promoter and 12-O-tetradecanoylphorbol-13-acetate-responsive elements. In a mutant mink lung epithelial cell line lacking T beta R-I, T beta R-I cDNA stimulated promoter activity, but the T beta R-1.2 cDNA did not. T beta R-2.1 formed an oligomer with T beta R-II when transfected into COS cells, but the complex did not transduce the signal after ligand stimulation. On the other hand, co-transfection of T beta R-1.2 and T beta R-2.1 cDNAs restored the responsiveness to TGF-beta 1. These results indicate that an interaction between the intracellular regions of T beta R-I and T beta R-II, triggered by ligand binding to the extracellular domains of these receptors, leads to efficient signal transduction by TGF-beta.

• Cusella-De Angelis MG et al.
• Differential response of embryonic and fetal myoblasts to TGF beta: a possible regulatory mechanism of skeletal muscle histogenesis.
• Development. 1994; 120: 925-33
• Display abstract

• Embryonic and fetal skeletal myoblasts were grown in culture in the presence of TGF beta. Under the conditions employed, TGF beta inhibited differentiation of fetal but not of embryonic myoblasts. To investigate the possible relevance of these data to skeletal muscle histogenesis in vivo, we studied the proliferation/differentiation state of mesodermal cells in the proximal region of the limb bud at the time of primary fiber formation. BrdU labeling and immunostaining for myosin heavy chains revealed that very few mesodermal cells enter the S phase of the cycle when differentiated primary fibers first appear. However, a few hours later, many cells in S phase surround newly formed muscle fibers, suggesting that the latter may be a source of mitogens for undifferentiated myoblasts. Co-culture experiments supported this hypothesis, showing that medium conditioned by fiber-containing explants can stimulate myoblast proliferation. Taken together these data suggested a possible mechanism for the regulation of muscle fiber formation. The model assumes that fibers form in the proximal region of the limb bud, where TGF beta is known to be present, and BrdU labeling experiments did not reveal cells in S phase. It is conceivable that non-dividing embryonic myoblasts (which do not respond to TGF beta) can undergo differentiation, while fetal myoblasts are inhibited by TGF beta. Once formed, primary fibers may stimulate a new wave of proliferation in fetal myoblasts, in order to expand the pool of cells needed to form secondary fibers.(ABSTRACT TRUNCATED AT 250 WORDS)

• Lopez-Casillas F, Payne HM, Andres JL, Massague J
• Betaglycan can act as a dual modulator of TGF-beta access to signaling receptors: mapping of ligand binding and GAG attachment sites.
• J Cell Biol. 1994; 124: 557-68
• Display abstract

• Betaglycan, also known as the TGF-beta type III receptor, is a membrane-anchored proteoglycan that presents TGF-beta to the type II signaling receptor, a transmembrane serine/threonine kinase. The betaglycan extracellular region, which can be shed by cells into the medium, contains a NH2-terminal domain related to endoglin and a COOH-terminal domain related to uromodulin, sperm receptors Zp2 and 3, and pancreatic secretory granule GP-2 protein. We identified residues Ser535 and Ser546 in the uromodulin-related region as the glycosaminoglycan (GAG) attachment sites. Their mutation to alanine prevents GAG attachment but does not interfere with betaglycan stability or ability to bind and present TGF-beta to receptor II. Using a panel of deletion mutants, we found that TGF-beta binds to the NH2-terminal endoglin-related region of betaglycan. The remainder of the extracellular domain and the cytoplasmic domain are not required for presentation of TGF-beta to receptor II; however, membrane anchorage is required. Soluble betaglycan can bind TGF-beta but does not enhance binding to membrane receptors. In fact, recombinant soluble betaglycan acts as potent inhibitor of TGF-beta binding to membrane receptors and blocks TGF-beta action, this effect being particularly pronounced with the TGF-beta 2 isoform. The results suggest that release of betaglycan into the medium converts this enhancer of TGF-beta action into a TGF-beta antagonist.

• Reisenbichler H, Jirtle RL
• BSA treatment of plasticware reduces TGF beta binding.
• Biotechniques. 1994; 17: 675-6

• Attisano L, Wrana JL, Lopez-Casillas F, Massague J
• TGF-beta receptors and actions.
• Biochim Biophys Acta. 1994; 1222: 71-80

• Hogan BL
• Developmental signalling. Sorting out the signals.
• Curr Biol. 1994; 4: 1122-4
• Display abstract

• New insights into the developmental roles played by the TGF-beta family of signalling molecules come from the identification in Drosophila of two transmembrane receptors encoded by the thick veins and saxophone genes.

• Bassing CH, Yingling JM, Wang XF
• Receptors for the TGF-beta ligand family.
• Vitam Horm. 1994; 48: 111-56

• Ueno N
• [Receptors for TGF-beta superfamily--a new signal transduction pathway of growth factor]
• Seikagaku. 1994; 66: 158-62

• Wall NA, Hogan BL
• TGF-beta related genes in development.
• Curr Opin Genet Dev. 1994; 4: 517-22
• Display abstract

• Embryonic induction is the process by which signals from one cell population change the developmental fate of another. Polypeptides related to growth factors are one group of molecules mediating many inductive events. Recent data on the embryonic expression and function of signaling proteins related to transforming growth factor beta, in both vertebrate and invertebrate systems, have shown that these molecules play important roles in both pattern formation and tissue specification during embryogenesis.

• Filvaroff EH, Ebner R, Derynck R
• Inhibition of myogenic differentiation in myoblasts expressing a truncated type II TGF-beta receptor.
• Development. 1994; 120: 1085-95
• Display abstract

• Transforming growth factor-beta (TGF-beta) is thought to play a role in mesenchymal cell development and, specifically, in muscle differentiation, yet its precise role in the latter process remains unclear. TGF-beta has been shown to both inhibit and induce myoblast maturation in vitro, depending on the culture conditions. Whether the type I or type II TGF-beta receptor mediates the various TGF-beta effects on myogenesis is not known. In the present study, C2C12 myoblasts were transfected with an expression vector for a truncated type II TGF-beta receptor, which has been shown to act as a dominant negative inhibitor of type II receptor signaling. In contrast to the parental cells, the transfected clones did not efficiently form myotubes or induce expression of MyoD, myogenin and several other differentiation markers following incubation in low serum media. However, some muscle differentiation markers continued to be expressed in the transfected cells suggesting that at least two pathways are involved in muscle cell differentiation. These cells could still growth arrest in low serum media, showing that decreased proliferation can be dissociated from differentiation. Unlike several oncogenes known to block myogenic differentiation, expression of the truncated TGF-beta receptor did not result in myoblast transformation. Injection of the parental or the transfected C2C12 cells into the limb muscle of nude mice revealed quantitative and qualitative differences in their behavior, and suggested that myoblasts expressing the truncated TGF-beta receptor cannot fuse in vivo. Finally, retrovirus-mediated expression of MyoD in the transfected cells restored their ability to form myotubes in vitro, indicating that inhibition of myoblast differentiation by the truncated TGF-beta receptor may depend on decreased MyoD expression. We propose that TGF-beta signaling through the type II receptor is required for several distinct aspects of myogenic differentiation and that TGF-beta acts as a competence factor in this multistep process.

• Jaskoll T, Choy HA, Melnick M
• Glucocorticoids, TGF-beta, and embryonic mouse salivary gland morphogenesis.
• J Craniofac Genet Dev Biol. 1994; 14: 217-30
• Display abstract

• Branching morphogenesis is a fundamental embryologic process in many developing organs: salivary gland, lung, pancreas, mammary gland, and kidney. Glucocorticoids (CORT) are known to regulate morphogenesis in several branching organs (e.g., lung); we hypothesize that CORT is also important to embryonic mouse salivary gland morphogenesis. We studied the CORT-glucocorticoid receptor (GR) signal transduction pathway during embryonic mouse submandibular gland development. Western analysis demonstrates that the 96-kDa GR is present in E14 to E18 submandibular glands. The embryonic GR is functional, as defined by its ability to bind a DNA CORT response element, for all gestational ages evaluated (E14 to E18); increasing GR levels are observed with progressive development. The level of endogenous corticosterone in embryonic submandibular glands was also determined. Using radioimmunoassays corticosterone is first detected on day 15 of gestation; there is a progressive increase in steroid levels from day 15 to 18 of gestation. In addition, we investigated the effect of exogenous CORT on submandibular gland morphogenesis in vivo and in vitro. In vivo experiments were performed using maternal injection of CORT or sham injection on day 12 of gestation with sacrifice 72 hr post-injection (E15). CORT treatment significantly increases embryonic submandibular gland growth in vivo. To further analyze the effect of exogenous CORT on embryonic submandibular gland morphogenesis, E13 mouse submandibular glands were cultured under serum-free, chemically-defined conditions. A significant (P < 0.05) enhancement of mean branching ratios (72 hr/0 hr) is detected in CORT-supplemented explants (10(-5) M to 10(-8) M) compared to control explants; doses more dilute than 10(-8) M CORT do not significantly (P > 0.10) increase the induction of branching. In addition, CORT administration to embryonic mice in utero enhances the expression of acinar-cell-specific mucin protein. These data indicate that the CORT-GR signal transduction pathway plays an important role in salivary gland morphogenesis. Northern analysis of TGF-beta 1, TGF-beta 2, and TGF-beta 3 transcript levels in vivo confirms the presence of TGF-beta 1 and TGF-beta 2 and identifies TGF-beta 3 in embryonic submandibular glands. To begin to identify a molecular mechanism of CORT-GR mediated submandibular gland morphodifferentiation, we studied the effect of CORT on growth factor gene expression, specifically TGF-beta. Northern analysis suggests that the CORT-GR signal transduction pathway modulates the rate of morphogenesis by regulating TGF-beta 2 and TGF-beta 3 mRNA expression.

• Stambuk N
• TGF-beta-induced cataract-like changes in lens epithelia.
• Invest Ophthalmol Vis Sci. 1994; 35: 3787-8

• Coussens LM, Yokoyama K, Chiu R
• Transforming growth factor beta 1-mediated induction of junB is selectively inhibited by expression of Ad.12-E1A.
• J Cell Physiol. 1994; 160: 435-44
• Display abstract

• Transforming growth factor beta (TGF-beta), a multifunctional polypeptide growth factor, regulates the expression of many genes critical to cell cycle progression, such as members of the jun gene family which encode components of the transcription factor complex AP-1. The transforming proteins encoded by the early region 1A of adenovirus12 (Ad.12-E1A) abrogate some of the cellular responses to TGF-beta as well as affecting, differentially, the expression of cellular jun genes. Our data demonstrate that expression of Ad.12-E1A in rat 3Y1 fibroblast cells inhibits induction of junB by TGF-beta 1 while not altering the regulation of junB by phorbol ester or serum. Regulation of c-jun gene expression by TGF-beta 1, phorbol ester, and serum is not appreciably altered by the expression of Ad.12-E1A. Inhibition of TGF-beta induced junB expression is not due to a defect in TGF-beta/receptor interaction on Ad.12-E1A transformed cells and is not observed in other isotypic fibroblast cells transformed by SV40 or polyomavirus. These data suggest that multiple, independent, intracellular signal transduction pathways exist which mediate genomic responses to TGF-beta. Cellular expression of Ad.12-E1A-12S gene products results in selective disruption of some TGF-beta 1 signaling cascades and not those activated by phorbol ester or serum. These data further suggest that some cellular targets which mediate TGF-beta 1 action may also be unique targets of action for the E1A-12S transforming protein of adenovirus12.

• Xie T, Finelli AL, Padgett RW
• The Drosophila saxophone gene: a serine-threonine kinase receptor of the TGF-beta superfamily.
• Science. 1994; 263: 1756-9
• Display abstract

• The Drosophila decapentaplegic (dpp) gene encodes a transforming growth factor-beta (TGF-beta)-like protein that plays a key role in several aspects of development. Transduction of the DPP signal was investigated by cloning of serine-threonine kinase transmembrane receptors from Drosophila because this type of receptor is specific for the TGF-beta-like ligands. Here evidence is provided demonstrating that the Drosophila saxophone (sax) gene, a previously identified female sterile locus, encodes a TGF-beta-like type I receptor. Embryos from sax mothers and dpp embryos exhibit similar mutant phenotypes during early gastrulation, and these two loci exhibit genetic interactions, which suggest that they are utilized in the same pathway. These data suggest that sax encodes a receptor for dpp.

• Lin HY, Moustakas A
• TGF-beta receptors: structure and function.
• Cell Mol Biol (Noisy-le-grand). 1994; 40: 337-49
• Display abstract

• The transforming growth factor beta (TGF-beta) superfamily of dimeric polypeptide growth factors regulate cell growth and differentiation and play important roles in embryonic development and the immune response. The molecular cloning of three types of high affinity cell-surface receptors (type I, type II and type III) for these growth factors provide insights into their poorly understood mechanisms of action. Here we focus on the structure of these receptors and on the complex interactions which occur between different receptors that are necessary for ligand-induced signal transduction. The type I and type II receptors are both related transmembrane serine/threonine kinase receptors which may hetero-oligomerize to transduce proper signals. The type III receptor modulates the binding of ligands to the signaling complex of type I and type II receptors. These findings suggest that serine/threonine phosphorylation and heteromeric receptor interactions are important elements of TGF-beta induced signaling.

• Derynck R
• TGF-beta-receptor-mediated signaling.
• Trends Biochem Sci. 1994; 19: 548-53
• Display abstract

• Transforming growth factor beta (TGF-beta) and its many relatives are thought to play key roles in the control of cell proliferation and differentiation. In particular, the ability of TGF-beta to induce growth arrest in epithelial cells has drawn considerable attention. The recent cloning of TGF-beta receptors, which are considered to be prototypes of a new class of cell-surface receptors, has provided a first insight into how TGF-beta signaling induces a variety of intracellular changes. Furthermore, recent advances in the characterization of the cell-cycle machinery have stimulated studies aimed at understanding how TGF-beta signaling leads to growth arrest in the late G1 phase of the cell cycle.

• Kogawa K, Mogi Y, Morii K, Takimoto R, Niitsu Y
• [TGF-beta and platelet]
• Rinsho Ketsueki. 1994; 35: 370-5
• Display abstract

• In the present paper, we investigated the pathophysiological implication of TGF-beta from megakaryocytes or megakaryoblasts in the development of myelofibrosis. In the bone marrow of myelofibrosis, proliferation of megakaryocytes is often noticed. We therefore investigated the TGF-beta expression in the bone marrow megakaryocytes from 12 chronic myeloproliferative disorder patients with myelofibrosis by immunohistochemical analysis. About all the specimen showed strong positivity for TGF-beta. In order to examine whether megakaryoblasts produce TGF-beta, we then measured TGF-beta activity in the conditioned medium (CM) of megakaryoblasts from a patient with acute megakaryoblastic leukemia who had profound myelofibrosis. The CM showed strong collagen synthesis stimulating activity which was nullified by addition of anti TGF-beta antibody. Since TGF-beta exists as latent form in platelets, TGF-beta was considered to be altered from active to latent form during megakaryocytes differentiation. In this context, MEG-01, a megakaryoblastic cell line which produces active TGF-beta was underwent differentiation to produce platelet-like bleb with TPA treatment. During the differentiation, MEG-01 showed the decrease of active TGF-beta production and increase of latent TGF-beta together with the production of LTBP. These results suggest that megakaryoblasts produce active TGF-beta and may may cause myelofibrosis, while more differentiated megakaryocytes produce latent TGF-beta. Mechanism by which megakaryoblast escape from negative autocrine of active TGF-beta was also investigated. MEG-01 was found to express mutated p53 which is considered to be responsible for impaired signal transduction of TGF-beta.

• Burt DW, Law AS
• Evolution of the transforming growth factor-beta superfamily.
• Prog Growth Factor Res. 1994; 5: 99-118
• Display abstract

• Transforming growth factor beta 1 (TGF-beta 1) is the prototype of an increasingly complex superfamily of growth and differentiation factors. To date, a total of 74 TGF-beta-like sequences have been published, probably representing 23 distinct genes. These sequences were obtained from mammalian, avian, amphibian and insect species, thus emphasising the ancient nature of the TGF-beta superfamily peptides. This article summarises current hypotheses concerning the evolutionary history of this protein superfamily, based on the molecular phylogeny of the published sequences. Comparison of the deduced amino acid sequences leads to the definition of five main groups within the superfamily (TGF-beta, Bone Morphogenetic Proteins [BMP], Anti-Mullerian Hormone [AMH], Inhibin alpha [INH alpha] and GDF-9) and six subgroups within the BMPs (60A, Decapentaplegic [dpp], Vg1, BMP-3, Inhibin beta [INH beta A/B] and nodal). This classification predicts possible phylogenetic and functional relationships among these proteins.

• Kingsley DM
• The TGF-beta superfamily: new members, new receptors, and new genetic tests of function in different organisms.
• Genes Dev. 1994; 8: 133-46

• Pawson T
• Introduction: protein kinases.
• FASEB J. 1994; 8: 1112-3

• Yamashita H, Ichijo H, Grimsby S, Moren A, ten Dijke P, Miyazono K
• Endoglin forms a heteromeric complex with the signaling receptors for transforming growth factor-beta.
• J Biol Chem. 1994; 269: 1995-2001
• Display abstract

• Human endoglin is a dimeric protein that binds transforming growth factor-beta (TGF-beta). A porcine cDNA clone for endoglin was obtained from a porcine uterus cDNA library. The deduced sequence of the primary translated product of endoglin consists of 643 amino acids with a high sequence identity (96%) to human endoglin in the transmembrane and intracellular domains, but with a lower sequence similarity (66%) in the extracellular domain. In contrast to human endoglin, porcine endoglin has no Arg-Gly-Asp tripeptide in its sequence. Antibodies, raised against a peptide corresponding to the intracellular domain of porcine endoglin, immunoprecipitated an 84-kDa protein under reducing condition and a 130-kDa protein under nonreducing condition in porcine aortic endothelial cells. Porcine endoglin bound TGF-beta 1 and -beta 3 efficiently, but TGF-beta 2 less efficiently. Endoglin was found to be coimmunoprecipitated with TGF-beta receptors type I and/or II by the endoglin antibodies or by TGF-beta receptor II antibodies in the presence of ligand. Thus, endoglin and TGF-beta receptors I and/or II most likely formed a heteromeric receptor complex. Endoglin was phosphorylated on serine residue(s), which did not change after stimulation by TGF-beta 1. These results revealed that endoglin is a phosphorylated protein which forms a heteromeric complex with signaling receptors for TGF-beta.

• Storm EE, Huynh TV, Copeland NG, Jenkins NA, Kingsley DM, Lee SJ
• Limb alterations in brachypodism mice due to mutations in a new member of the TGF beta-superfamily.
• Nature. 1994; 368: 639-43
• Display abstract

• The mutation brachypodism (bp) alters the length and number of bones in the limbs of mice but spares the axial skeleton. It illustrates the importance of specific genes in controlling the morphogenesis of individual skeletal elements in the tetrapod limb. We now report the isolation of three new members of the transforming growth factor-beta (TGF-beta) superfamily (growth/differentiation factors (GDF) 5,6 and 7) and show by mapping, expression patterns and sequencing that mutations in Gdf5 are responsible for skeletal alterations in bp mice. GDF5 and the closely related GDF6 and GDF7 define a new subgroup of factors related to known bone- and cartilage-inducing molecules, the bone morphogenetic proteins (BMPs). Studies of Bmp5 mutations in short ear mice have shown that at least one other BMP gene is also required for normal skeletal development. The highly specific skeletal alterations in bp and short ear mice suggest that different members of the BMP family control the formation of different morphological features in the mammalian skeleton.

• Tokuyama Y, Tokuyama H
• Purification and identification of TGF-beta 2-related growth factor from bovine colostrum.
• J Dairy Res. 1993; 60: 99-109
• Display abstract

• Bovine colostrum contains transforming growth factor (TGF)-beta-like activity. High levels of this activity are found in early colostrum (within 12 h after parturition); however, it decreases rapidly after 30 h. In this study, the activity in early colostrum was purified by a sequence of decaseination, DEAE-Sephacel chromatography, Sephadex G-100 gel filtration, precipitation at neutral pH, further gel filtration and reversed-phase FPLC. A homodimeric 23 kDa protein was identified that retained the TGF-beta-like activity. Its activity was completely neutralized by a specific anti-TGF-beta 2 antibody. These results indicate that the TGF-beta-like growth factor found in bovine colostrum is chemically and antigenically related to TGF-beta 2. The physiological roles of the TGF-beta 2-related growth factor are discussed.

• Blessing M, Nanney LB, King LE, Jones CM, Hogan BL
• Transgenic mice as a model to study the role of TGF-beta-related molecules in hair follicles.
• Genes Dev. 1993; 7: 204-15
• Display abstract

• There is increasing evidence that members of the TGF-beta superfamily are important regulators of epithelial growth and differentiation in vivo. Here, transgenic mice have been used to study the role of the TGF-beta-related growth factors BMP-2 and BMP-4 in hair and whisker development. In the mature hair follicle, BMP-2 transcripts are normally seen only in precortex cells at the base of the hair shaft. In the transgenic mice reported here, BMP-4, a closely related molecule, has been ectopically expressed in the outer root sheath of hair and whisker follicles using an expression vector based on the bovine cytokeratin IV* promoter. In response to transgene expression, both outer root sheath cells below the stem cell compartment and hair matrix cells around the dermal papilla cease proliferation. In addition, the expression pattern of cytokeratin markers is disturbed in some transgenic hair follicles. These results support a model in which members of the TGF-beta superfamily play an active role in the inhibiton of cell proliferation and the onset of expression of trichocyte-specific genes that take place when cells leave the matrix of the follicle and differentiate into shaft cells.

• Laurent GJ, Coker RK, McAnulty RJ
• TGF-beta antibodies: a novel treatment for pulmonary fibrosis?
• Thorax. 1993; 48: 953-4

• Daopin S, Li M, Davies DR
• Crystal structure of TGF-beta 2 refined at 1.8 A resolution.
• Proteins. 1993; 17: 176-92
• Display abstract

• The crystal structure of TGF-beta 2 has been refined using data collected with synchrotron radiation (CHESS) to 1.8 A resolution with a residual R (= sigma magnitude of Fo-magnitude of Fc/sigma magnitude of Fo) factor of 17.3%. The model consists of 890 protein atoms from all 112 residues and 59 water molecules. The monomer of TGF-beta 2 assumes a rather extended conformation and lacks a well-defined hydrophobic core. Surface accessibility calculations show only 44% of the nonpolar surface is buried in the monomer. In contrast, 55.8% of the nonpolar surface area is buried when the two monomers form a dimer, a typical value for globular proteins. This includes a 1300 A2 buried interface area that is largely hydrophobic. Sequence comparisons using a profile derived from the refined TGF-beta 2 structure suggest that the cluster of four disulfides (three intramonomeric disulfide bonds 15-78, 44-109, 48-111 forming a disulfide knot, and one intermonomeric disulfide 77-77) together with the extended beta strand region constitutes the conserved structural motif for the TGF-beta superfamily. This structural motif, without the 77-77 disulfide bond, defines also the common fold for a general family of growth factors, including the nerve growth factor and platelet-derived growth factor families. The fold is conserved only at the monomer level, while the active forms are dimers, suggesting that dimerization plays an important role in regulating the binding of these cytokines to their receptors and in modulating the biological responses.

• MacLellan WR, Brand T, Schneider MD
• Transforming growth factor-beta in cardiac ontogeny and adaptation.
• Circ Res. 1993; 73: 783-91
• Display abstract

• The transforming growth factor-beta (TGF-beta) superfamily comprises a set of regulatory peptides with multiple effects on cell growth and differentiation. The elaborate regulation of TGF-beta s during embryonic development of the heart, the upregulation of TGF-beta after hemodynamic stress, and the impact of TGF-beta on cardiac gene expression together imply a prominent functional role for this family of growth factors in cardiac organogenesis and hypertrophy. Basal and TGF-beta-induced expression of skeletal alpha-actin, one of several genes specifically associated with developing or hypertrophied myocardium, each are contingent on transcriptional activation by serum response factor. A truncated form of the type II TGF-beta receptor, created by deletion of the cytoplasmic kinase domain, acts as a dominant suppressor of TGF-beta signal transduction in cultured cardiac muscle cells and may provide a suitable means to establish the functions of TGF-beta in vivo.

• Lahm H, Odartchenko N
• Role of transforming growth factor beta in colorectal cancer.
• Growth Factors. 1993; 9: 1-9

• Pronost S, Redini F, Vivien D, Galera P, Pujol JP
• Human rheumatoid synovial cells (HRSC) in culture express TGF-beta receptors and are growth stimulated by the factor.
• Agents Actions Suppl. 1993; 39: 133-7
• Display abstract

• TGF-beta (5 ng/ml) increased the proliferation rate of HRSC by 30% after 96 h of treatment. One type of TGF-beta binding system, with apparent Kd of 953 pM and a number of 29,400 receptors/cell, was detected by Scatchard analysis of [125I]-TGF-beta binding. However, crosslinking experiments and SDS-PAGE separation showed five TGF-beta binding proteins: 50, 70, 110, 140 and 400 kDa. We may suggest that the affinities of these receptors are too close to be revealed by Scatchard plot. All together, the data suggest that TGF-beta plays a role in the hyperplasia of RA synovial tissue.

• Lopez-Casillas F, Wrana JL, Massague J
• Betaglycan presents ligand to the TGF beta signaling receptor.
• Cell. 1993; 73: 1435-44
• Display abstract

• Transforming growth factor beta (TGF beta) signals through a heteromeric protein kinase receptor that has a limited ability to bind ligand. This limitation is overcome by the action of betaglycan (TGF beta type III receptor), a separate TGF beta-binding membrane protein of previously unknown function. Betaglycan presents TGF beta directly to the kinase subunit of the signaling receptor, forming a high affinity ternary complex. Membrane betaglycan increases TGF beta binding to the signaling receptor, enhances cell responsiveness to TGF beta, and eliminates marked biological differences between TGF beta isoforms. Thus, betaglycan is a direct regulator of TGF beta access to the signaling receptors.

• Macias D, Ganan Y, Hurle JM
• Modification of the phalangeal pattern of the digits in the chick embryo leg bud by local microinjection of RA, staurosporin and TGF beta's.
• Anat Embryol (Berl). 1993; 188: 201-8
• Display abstract

• Many experimental studies show that in the avian chick limb the digits are specified at early stages of development by characteristic concentrations within the limb mesoderm of a still unidentified morphogen diffusing from the posterior margin of the bud, linked with a specific pattern of homeobox gene expression. In all these studies, digits are distinguished by their size, morphology and phalangeal pattern rather than by their position within the autopodium. In this work we report the induction of digits that have otherwise normal morphology but lack an interphalangeal joint. This suggests that the patterning of these joints is not necessarily linked to the control of the outgrowth of the digital rays. Missing interphalangeal joints were induced by microinjection into the third interdigital space of the leg bud of stage 28 to 31 chick embryos of retinoic Acid (RA), staurosporine and TGF beta 1 and beta 2, but not by microinjection of FGF or EGF. Our results also suggest that the pattern of insertion of the long tendons and the formation of the flexor cutaneous pad at the plantar surface of the digits are both linked to the establishment of the interphalangeal joints.

• Luo LD, Wirth PJ
• Consecutive silver staining and autoradiography of 35S and 32P-labeled cellular proteins: application for the analysis of signal transducing pathways.
• Electrophoresis. 1993; 14: 127-36
• Display abstract

• The methodology for the simultaneous analysis of protein synthesis concomitant with protein phosphorylation/dephosphorylation is described. The technique consists of metabolic labeling of rat liver epithelial (RLE) cells with [32P]orthophosphate and [35S]methionine, performing two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) of the mixed samples, followed by silver staining and subsequent autoradiography of the dried silver stained 2-D PAGE electrophoretograms using two films placed back-to-back. The first film, which is positioned in direct contact with the dried silver-stained gel, visualized both exposure to 35S and 32P while the second film recorded exposure to only 32P due to the differential energy levels of the two isotopes. The juxta-positioning of the silver-stained images with the two autoradiographic film images permits the unambiguous mapping of the phosphorylated polypeptides back to their corresponding silver-stained and methionine-labeled counterparts. This strategy provides quantitative information utilizing both silver staining (measure of constitutive levels of protein expression) and metabolic labeling to measure rates of protein synthesis and/or degradation and phosphorylation and/or dephosphorylation using [35S]methionine and [32P]orthophosphate, respectively. We have utilized this methodology for the in vitro analysis of transforming growth factor type beta 1 (TGF-beta 1)-mediated signal transduction in RLE cells and have identified three nuclear polypeptides, 1 (pI 4.95/M(r) 97 kDa), 2 (5.00/85 kDa) and 3 (4.90/84 kDa) whose phosphorylation status is rapidly and transiently modulated by TGF-beta 1. The methodology described should have wide applications in studies where it is desirous to measure protein synthesis and/or degradation concomitant with signal transduction pathways involving protein phosphorylation.

• Mukai M, Shinkai K, Yoshioka K, Imamura F, Akedo H
• [Mechanism of tumor cell invasion studied by a culture model--modification of invasiveness by host mediators]
• Hum Cell. 1993; 6: 194-8
• Display abstract

• A culture model for invasion of rat mesothelial cell layer by rat ascites hepatoma cells has been developed. By using this quantitative model, we have recently found that the invasiveness of tumor cells is not only genetically determined but is greatly influenced by their interactions with host cells and host mediators. The preculture with macrophages was found to enhance both the in vitro and in vivo invasive potentials of the tumor cells. This potentiation appears to be mediated partly by oxygen radicals generated by the cocultured macrophages. The in vitro invasive capacity was also augmented by pretreating the tumor cells with TGF-beta, or with activated platelets. In the in vitro invasion assay system, tumor cells did not invade against cultured mesothelial cell monolayers without fetal calf serum. Serum could be completely substituted by oleoyl-lysophosphatidic acid (LPA) or bacterial phospholipase D (PLD), suggesting a possible participation of particular signaling cascade, PLD-LPA(PA) system, in the invasion of certain tumor cells.

• Vivien D et al.
• IPG (inositolphosphate glycan) as a cellular signal for TGF-beta 1 modulation of chondrocyte cell cycle.
• J Cell Physiol. 1993; 155: 437-44
• Display abstract

• The knowledge of transforming growth factor (TGF)-beta receptors has greatly progressed in the recent years. TGF-beta receptors type I and II have been implicated in the modulation of cell proliferation, whereas type III (betaglycan) may act as a component presenting TGF-beta to its signaling receptors. In addition, four other proteins that bind TGF-beta 1 or TGF-beta 2 have been recently identified in some cell lines, three being anchored to the membrane through a glycosylphosphatidylinositol (GPI). Despite this knowledge, the molecular mechanism of signal transduction through the TGF-beta receptors remain an enigma. TGF-beta family does not signal via any of the classical pathways. As GPI anchors of membrane proteins have been implicated in the transduction of some hormonal effects, we investigated the putative role of GPI in signaling the TGF-beta effects on the proliferation of rabbit articular chondrocytes (RAC). We previously showed that TGF-beta 1 increased DNA replication rate of RAC, with a recruitment of cells in G2/M followed by a subsequent mitosis wave. Here, we find that the factor causes specific GPI hydrolysis, with correlated increase of inositolphosphate glycan (IPG). This effect was specifically inhibited by antibodies that bind TGF-beta 1. Using [3H]-inositol labeling and Triton X-114 extraction, we demonstrate that a hydrophobic material from the membrane is cleaved by treatment of cell cultures with phosphatidylinositol specific phospholipase C (PI-PLC) or by exposure to TGF-beta, supporting that a PI-anchored molecule gives rise to IPG by TGF-beta-induced hydrolysis. The biological relevance of this hydrolysis was demonstrated by the enhancing effect of purified IPG on the DNA synthesis rate, which mimicked the TGF-beta action. These results demonstrate that IPG could be an early messenger in the cellular signaling that mediates the effect of TGF-beta on RAC growth.

• Hursh DA, Padgett RW, Gelbart WM
• Cross regulation of decapentaplegic and Ultrabithorax transcription in the embryonic visceral mesoderm of Drosophila.
• Development. 1993; 117: 1211-22
• Display abstract

• The Drosophila decapentaplegic gene (dpp) encodes a TGF-beta family member involved in signal transduction during embryonic midgut formation. The shortvein (shv) class of cis-regulatory dpp mutants disrupt expression in parasegments 4 and 7 (ps4 and ps7) of the embryonic visceral mesoderm (VM) surrounding the gut and cause abnormalities in gut morphogenesis. We demonstrate that cis-regulatory elements directing expression in ps4 and ps7 are separable and identify DNA fragments that generate ps4 and ps7 expression patterns using reporter gene constructs. dpp reporter gene expression in both ps4 and ps7 is autoregulated as it requires endogenous dpp+ activity. Reporter gene ps7 expression requires the wild-type action of Ultra-bithorax (Ubx), and abdominal-A. Furthermore, the expression of certain Ubx reporter genes is coincident with dpp in the VM. Both the mis-expression of Ubx reporter genes in the developing gastric caecae at ps4 and its normal expression in ps7 are dependent upon endogenous dpp+ activity. We conclude that dpp both responds to and regulates Ubx in ps7 of the visceral mesoderm and that Ubx autoregulation within this tissue may be indirect as it requires more components than have previously been thought.

• Segarini PR
• TGF-beta receptors: a complicated system of multiple binding proteins.
• Biochim Biophys Acta. 1993; 1155: 269-75

• Filmus J, Kerbel RS
• Development of resistance mechanisms to the growth-inhibitory effects of transforming growth factor-beta during tumor progression.
• Curr Opin Oncol. 1993; 5: 123-9
• Display abstract

• Members of the transforming growth factor-beta family, especially transforming growth factor-beta type 1, are among the most potent growth-inhibitory factors for epithelial, lymphohematopoietic, and neuroectodermal cells. Resistance to transforming growth factor-beta-mediated growth inhibition is frequently observed in cancers derived from these cell types. We review two important aspects of cancer cell resistance to transforming growth factor-beta: 1) It occurs progressively during the multistep process of tumor progression, and 2) it may encompass a potentially wide range of mechanisms involving transforming growth factor-beta-receptor alterations and cell-signaling defects. Stepwise increases in resistance to the growth-inhibitory action of transforming growth factor-beta may therefore sometimes be attributed to not one but several defects that are involved in transforming growth factor-beta activation, binding, and signaling. These factors accumulate within expanding tumor subclones during disease progression.

• Rijksen G, Voller MC, van Zoelen EJ
• The role of protein tyrosine phosphatases in density-dependent growth control of normal rat kidney cells.
• FEBS Lett. 1993; 322: 83-7
• Display abstract

• In normal rat kidney cells protein tyrosine phosphatases (PTPases) play a role in attaining density-dependent growth arrest after stimulation with mitogens. The PTPase inhibitor sodium orthovanadate prevents density-dependent growth inhibition of EGF-treated cells and mimicks in that respect the action of TGF beta and retinoic acid. However, enhanced PTPase activity is not obligatory for maintaining cells in a density-arrested state. In contrast to TGF beta and retinoic acid, vanadate is unable to restimulate density-inhibited cells, indicating that different mechanisms are operating. Yet, vanadate is strongly potentiating the effect of low concentrations of TGF beta but not of retinoic acid, implicating that tyrosine phosphorylation is linked to TGF beta action and that PTPase may represent a negative control element in the TGF beta signaling pathway.

• Lord E et al.
• Transfection of TGF-beta producing tumors with IL-2 elicits tumor rejection.
• Ann N Y Acad Sci. 1993; 690: 346-8

• Heberlein U, Wolff T, Rubin GM
• The TGF beta homolog dpp and the segment polarity gene hedgehog are required for propagation of a morphogenetic wave in the Drosophila retina.
• Cell. 1993; 75: 913-26
• Display abstract

• Development of the Drosophila retina occurs asynchronously; differentiation, its front marked by the morphogenetic furrow, progresses across the eye disc epithelium over a 2 day period. We have investigated the mechanism by which this front advances, and our results suggest that developing retinal cells drive the progression of morphogenesis utilizing the products of the hedgehog (hh) and decapentaplegic (dpp) genes. Analysis of hh and dpp genetic mosaics indicates that the products of these genes act as diffusible signals in this process. Expression of dpp in the morphogenetic furrow is closely correlated with the progression of the furrow under a variety of conditions. We show that hh, synthesized by differentiating cells, induces the expression of dpp, which appears to be a primary mediator of furrow movement.

• Roberts AB, Sporn MB
• Physiological actions and clinical applications of transforming growth factor-beta (TGF-beta).
• Growth Factors. 1993; 8: 1-9

• Rifkin DB, Kojima S, Abe M, Harpel JG
• TGF-beta: structure, function, and formation.
• Thromb Haemost. 1993; 70: 177-9

• Flaumenhaft R, Kojima S, Abe M, Rifkin DB
• Activation of latent transforming growth factor beta.
• Adv Pharmacol. 1993; 24: 51-76

• Nilsen-Hamilton M, Matrisian LM, Hamilton RT
• Developing in a family way. Transforming Growth Factor Type beta and Related Proteins in Development: the Sixth Molecular, Cellular and Developmental Biology/Iowa State University Symposium, Ames, IA, USA September 20-23, 1991.
• New Biol. 1992; 4: 127-31
• Display abstract

• We are left with the impression that members of the TGF-beta family are critically positioned in a cascade of events regulating complex developmental processes. Roberts described the TGF-betas as providing the cells with cues to their temporal positions in a developmental program, that is, telling the cells "where they were, where they are, and where they're going." The broad diversity of cellular and tissue responses to TGF-betas and the widespread expression of their receptors suggest the TGF-betas may act as a "common currency," enabling diverse cell types to communicate with each other. Other members of the family are more restricted in their expression and the expression of their receptors, and may have a more limited and well-defined developmental role. The complex regulation of the members of the TGF-beta family is consistent with their importance as directors and coordinators of complicated physiological processes such as those occurring in the development of multicellular organisms. Their expression is highly regulated at the transcriptional and post-transcriptional levels, and their localization and activation can be affected by binding proteins and matrix proteins. We still have much to learn about the receptors for this family of growth factors, but the recent cloning of the activin and TGF-beta receptors and the discovery of their enzymatic nature has dramatically opened the way for future studies to resolve the signal transduction pathway(s) used by members of this family.(ABSTRACT TRUNCATED AT 250 WORDS)

• Doctor JS, Jackson PD, Rashka KE, Visalli M, Hoffmann FM
• Sequence, biochemical characterization, and developmental expression of a new member of the TGF-beta superfamily in Drosophila melanogaster.
• Dev Biol. 1992; 151: 491-505
• Display abstract

• More than 20 members of the transforming growth factor-beta (TGF-beta) superfamily of growth and differentiation factors have been implicated in development. One member of the TGF-beta family has been previously reported from Drosophila, the decapentaplegic (dpp) gene which is involved in embryonic dorsal/ventral polarity, embryonic gut formation, and imaginal disk development. Using PCR methods, we have identified a second Drosophila gene in the TGF-beta family. It encodes a protein product that is more similar to the TGF-beta-related human bone morphogenetic proteins (BMPs) 5, 6, and 7 than it is to the Drosophila dpp gene product. Because of its localization on the polytene chromosome map, we refer to this gene as 60A. Expression of a 60A cDNA in Drosophila S2 cells was used to determine that 60A encodes a preproprotein that is processed to yield secreted amino- and carboxy-terminal polypeptides. The carboxy-terminal peptides are recovered as disulfide-linked homodimers. The 60A transcripts and protein are first detected at the onset of gastrulation, primarily in the mesoderm of the extending germ band. As the germ band retracts, and throughout later stages of embryonic development, the 60A transcript and protein are most readily detected in cells of the developing foregut and hindgut.

• Massague J, Weinberg RA
• Negative regulators of growth.
• Curr Opin Genet Dev. 1992; 2: 28-32
• Display abstract

• The proliferation of cells is regulated by countervailing positively- and negatively-acting signaling networks. The anti-proliferative signals, the study of which has been much neglected until recently, are often conveyed by growth-inhibitory peptides. Elements that mediate the cellular response to growth inhibitors are encoded by tumor suppressor genes that if lost may lead to the runaway growth of the cancer cell.

• Wrana JL et al.
• The type II TGF-beta receptor signals diverse responses in cooperation with the type I receptor.
• Cold Spring Harb Symp Quant Biol. 1992; 57: 81-6

• Hoffman M
• Researchers get a first look at the versatile TGF-beta family.
• Science. 1992; 257: 332-332

• Massague J
• Receptors for the TGF-beta family.
• Cell. 1992; 69: 1067-70

• Kramer IM, Koornneef I, de Laat SW, van den Eijnden-van Raaij AJ
• TGF-beta 1 induces phosphorylation of the cyclic AMP responsive element binding protein in ML-CCl64 cells.
• EMBO J. 1991; 10: 1083-9
• Display abstract

• Type beta transforming growth factors represent a family of polypeptides that modulate growth and differentiation. TGF-beta exerts its effects on target cells through interaction with specific cell surface receptors, but the signal transduction pathways are largely unresolved as yet. In this study we report that TGF-beta 1 induces a rapid phosphorylation of the cyclic AMP responsive element binding protein (CREB) in mink lung CCl64 cells. Phosphorylation induced by TGF-beta 1 is not mediated by the cAMP-dependent protein kinase. Parallel to the increase in phosphorylation of CREB, an increase in binding to the collagenase TPA responsive element was observed. CREB participates in the binding to this element, probably as a heterodimer with another as yet unknown protein. The modification imposed on CREB and its involvement in an enhanced TRE-binding could be a mechanism by which TGF-beta 1 induces the TRE-mediated transcriptional activation.

• Segarini PR
• TGF-beta receptors.
• Ciba Found Symp. 1991; 157: 29-40
• Display abstract

• TGF-beta binds specifically and with high affinity to all cell types with few exceptions. The binding parameters vary because the dissociation constants range from 1 pM to 60 pM. There may be as few as 200 or as many as 100,000 receptors per cell, depending on the cell type. Several cell surface proteins that specifically bind TGF-beta 1 and TGF-beta 2 have been characterized as putative receptors and have been classified on the basis of their size. Type I receptors have an Mr of approximately 50,000, type II receptors an Mr of approximately 80,000. Both type I and II receptors are glycoproteins and TGF-beta-induced responses appear to be mediated by one or both types. The most abundant and largest of the cell surface TGF-beta binding proteins is a membrane-bound proteoglycan that is a dimer of subunits each with Mr of approximately 250,000 (beta-glycan). This protein is plentiful on primary cells of mesenchymal origin but absent on primary epithelial and endothelial cells. It does not appear to be associated with TGF-beta-mediated cell responses and its function remains unknown. There are multiple cellular responses to TGF-beta and accordingly there appear to be multiple pathways for signal transduction. A guanine nucleotide binding protein-dependent pathway is involved in transmission of the signal for at least one TGF-beta-induced response while there is evidence that other responses are mediated through an independent pathway. TGF-beta receptor purification and cloning efforts will be rewarded with valuable information on the mechanisms of signal transmission.

• Massague J, Cheifetz S, Boyd FT, Andres JL
• TGF-beta receptors and TGF-beta binding proteoglycans: recent progress in identifying their functional properties.
• Ann N Y Acad Sci. 1990; 593: 59-72

• Sporn MB, Roberts AB
• TGF-beta: problems and prospects.
• Cell Regul. 1990; 1: 875-82
• Display abstract

• TGF-beta research is proceeding at an exponential pace. Studies in this area have become increasingly relevant to many areas of cell regulation, continually providing new surprises and findings. One may confidently predict that TGF-beta will be one of the key molecules in future attempts to establish an integrated view of the processes whereby cells coordinate their own and mutual functions. We have seen only the beginning, and this minireview is only a glimpse.

• Moses HL, Yang EY, Pietenpol JA
• TGF-beta stimulation and inhibition of cell proliferation: new mechanistic insights.
• Cell. 1990; 63: 245-7

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