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JOURNAL ARTICLES
Cellular interpretation of multiple TGF-beta signals: intracellular antagonism between activin/BVg1 and BMP-2/4 signaling mediated by Smads
A.F. Candia, T. Watabe, S.H. Hawley, D. Onichtchouk, Y. Zhang, R. Derynck, C. Niehrs, K.W. Cho
Development 1997 124: 4467-4480;

Summary

During early embryogenesis of Xenopus, dorsoventral polarity of the mesoderm is established by dorsalizing and ventralizing agents, which are presumably mediated by the activity of an activin/BVg1-like protein and Bone Morphogenetic Proteins (BMP), respectively. Interestingly, these two TGF-beta subfamilies are found in overlapping regions during mesoderm patterning. This raises the question of how the presumptive mesodermal cells recognize the multiple TGF-beta signals and differentially interpret this information to assign a particular cell fate. In this study, we have exploited the well characterized model of Xenopus mesoderm induction to determine the intracellular interactions between BMP-2/4 and activin/BVg1 signaling cascades. Using a constitutively active BMP-2/4 receptor that transduces BMP-2/4 signals in a ligand-independent fashion, we demonstrate that signals provided by activin/BVg1 and BMP modulate each other's activity and that this crosstalk occurs through intracellular mechanisms. In assays using BMP-2/4 and activin/BVg1-specific reporters, we determined that the specificity of BMP-2/4 and activin/BVg1 signaling is mediated by Smad1 and Smad2, respectively. These Smads should be considered as the mediators of the intracellular antagonism between BMP-2/4 and activin/BVg1 signaling possibly through sequestration of a limited pool of Smad4. Consistent with such a mechanism, Smad4 interacts functionally with both Smad1 and −2 to potentiate their signaling activities, and a dominant negative variant of Smad4 can inhibit both activin/BVg1 and BMP-2/4 mediated signaling Finally, we demonstrate that an activin/BVg1-dependent transcriptional complex contains both Smad2 and Smad4 and thereby provides a physical basis for the functional involvement of both Smads in TGF-beta-dependent transcriptional regulation. Thus, Smad4 plays a central role in synergistically activating activin/BVg1 and BMP-dependent transcription and functions as an intracellular sensor for TGF-beta-related signals.

REFERENCES

    1. Baker J. C.,
    2. Harland R. M.
    (1996) A novel mesoderm inducer, Madr2, functions in the activin signal transduction pathway. Genes Dev 10, 1880–1889
    OpenUrlAbstract/FREE Full Text
    1. Blitz I. L.,
    2. Cho K. W. Y.
    (1995) Anterior neuroectoderm is progressively induced during gastrulation: the role of the Xenopus homeobox gene orthodenticle. Development 121, 993–1004
    OpenUrlAbstract
    1. Brummel T. J.,
    2. Twombly V.,
    3. Marques G.,
    4. Wrana J. L.,
    5. Newfeld S. J.,
    6. Attisano L.,
    7. Massague J.,
    8. O'Connor M. B.,
    9. Gelbart W. M.
    (1994) Characterization and relationship of Dpp receptors encoded by the saxophone and thick veins genes in Drosophila. Cell 78, 251–261
    OpenUrlCrossRefPubMedWeb of Science
    1. Chen X.,
    2. Rubock M. J.,
    3. Whitman M.
    (1996) A transcriptinal partner for MAD proteins in TGF-singalling. Nature 383, 691–696
    OpenUrlCrossRefPubMedWeb of Science
    1. Cho K. W. Y.,
    2. Blumberg G.,
    3. Steinbeisser H.,
    4. De Robertis E. M.
    (1991) Molecular nature of Spemann's organizer: the role of the Xenopus homeobox gene goosecoid. Cell 67, 1111–1120
    OpenUrlCrossRefPubMedWeb of Science
    1. Chomzynski P.,
    2. Sacchi N.
    (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analyt. Biochem 162, 156–159
    OpenUrlCrossRefPubMedWeb of Science
    1. Dale L.,
    2. Howes G.,
    3. Price B. M. J.,
    4. Smith J. C.
    (1992) Bone morphogenetic protein 4: a ventralizing factor in early Xenopus development. Development 115, 573–585
    OpenUrlAbstract
    1. Derynck R.,
    2. Zhang Y.
    (1996) The Mad way to do it. Curr. Biol 6, 1226–1229
    OpenUrlCrossRefPubMedWeb of Science
    1. de Wet, J. R.,
    2. Wood K. V.,
    3. DeLuca M.,
    4. Helinski D. R.,
    5. Subramani S.
    (1987) Firefly, luciferase gene: Structure and expression in mammalian cells. Mol. Cell Biol 7, 725–737
    OpenUrlAbstract/FREE Full Text
    1. Eppert K.,
    2. Scherer S. W.,
    3. Ozcelik H.,
    4. Pirone R.,
    5. Hoodless P.,
    6. Kim H.,
    7. Tsui L.-C.,
    8. Bapat B.,
    9. Gallinger S.,
    10. Andrulis I.,
    11. Thomson J. H.,
    12. Wrana J.,
    13. Attisano L.
    (1996) MADR2 maps to 18q21 and encodes a TGF--regulated MAD-related protein that is functionally mutated in colorectal carcinoma. Cell 86, 543–552
    OpenUrlCrossRefPubMedWeb of Science
    1. Fainsod A.,
    2. Steinbeisser H.,
    3. De Robertis E. M.
    (1994) On the function of BMP-4 in patterning the marginal zone of the Xenopus embryo. EMBO J 13, 5015–5025
    OpenUrlPubMedWeb of Science
    1. Gawantka V.,
    2. Delius H.,
    3. Hirschfeld K.,
    4. Blumenstock C.,
    5. Nihers C.
    (1995) Antagonizing the Spemann organizer: role of the homeobox gene Xvent-1. EMBO J 14, 6268–6279
    OpenUrlPubMedWeb of Science
    1. Graff J.,
    2. Thies R. S.,
    3. Song J. J.,
    4. Celeste A. J.,
    5. Melton D. A.
    (1994) Studies with a Xenopus BMP receptor suggest that ventral mesoderm-inducing signals override dorsal signals in vivo. Cell 79, 169–179
    OpenUrlCrossRefPubMedWeb of Science
    1. Graff J. M.,
    2. Bansal A.,
    3. Melton D. A.
    (1996) Xenopus mad proteins transduce distinct subsets of signals for the TGFsuperfamily. Cell 85, 479–487
    OpenUrlCrossRefPubMedWeb of Science
    1. Green J. B.,
    2. Smith J. C.
    (1990) Graded changes in dose of a Xenopus activin A homologue elicit stepwise transitions in embryonic cell fate. Nature 347, 391–394
    OpenUrlCrossRefPubMed
    1. Hawley S. H. B.,
    2. Wunnenberg-Stapleton K.,
    3. Hashimoto C.,
    4. Laurent M. N.,
    5. Watabe T.,
    6. Blumberg B. W.,
    7. Cho K. W. Y.
    (1995) Disruption of BMP signals in embryonic Xenopus ectoderm leads to direct neural induction. Genes Dev 9, 2923–2935
    OpenUrlAbstract/FREE Full Text
    1. Hogan B. L.
    (1996) Bone morphogenetic proteins: multifunctinal regulators of vertebrate development. Genes Dev 10, 1580–1594
    OpenUrlFREE Full Text
    1. Hoodless P. A.,
    2. Haerry T.,
    3. Abdollah S.,
    4. Stapleton M.,
    5. O'Connor M. B.,
    6. Attisano L.,
    7. Wrana J. L.
    (1996) MADR1, a MAD-related protein that functions in BMP2 signaling pathways. Cell 85, 489–500
    OpenUrlCrossRefPubMedWeb of Science
    1. Howard E. W.,
    2. Knauer D. J.
    (1986) Biosynthesis of protease nexin-I. J. Biol. Chem 261, 14184–14190
    OpenUrl
    1. Jones C. M.,
    2. Dale L.,
    3. Hogan B. L. M.,
    4. Wright C. V. E.,
    5. Smith J. C.
    (1996) Bone morphogenetic protein-4 (BMP-4) acts during gastrula stages to cause ventralization of Xenopus embryos. Development 122, 1545–1554
    OpenUrlAbstract
    1. Jones C. M.,
    2. Kuehn M. R.,
    3. Hogan B. L.,
    4. Smith J. C.,
    5. Wright C. V.
    (1995) Nodal-related signals induce axial mesoderm and dorsalize mesoderm during gastrulation. Development 121, 3651–62
    OpenUrlAbstract
    1. Jones C. M.,
    2. Lyons K. M.,
    3. Lapan P. M.,
    4. Wright C. V. E.,
    5. Hogan B. L. M.
    (1992) DVR-4 (bone morphogenetic protein-4) as a posterior-ventralizing factor in Xenopus mesoderm induction. Development 115, 639–647
    OpenUrlAbstract
    1. Kessler D. S.,
    2. Melton D. A.
    (1995) Induction of dorsal mesoderm by soluble, mature Vg1 protein. Development 121, 2155–64
    OpenUrlAbstract
    1. Kim J.,
    2. Johnson K.,
    3. Chen H.J.,
    4. Carroll S.,
    5. Laughon A.
    (1997) Drosophila MAD binds to DMA and directly mediates activation of vestigial by decapentaplegic. Nature 388, 304–308
    OpenUrlCrossRefPubMedWeb of Science
    1. Krieg P. A.,
    2. Melton D. A.
    (1984) Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucl. Acid Res 12, 7057–7070
    OpenUrlAbstract/FREE Full Text
    1. Lagna G.,
    2. Hata A.,
    3. Hemmati-Brivanlou A.,
    4. Massague J.
    (1996) Partnership between DPC4 and SMADproteins in TGF-signalling pathways. Nature 383, 832–836
    OpenUrlCrossRefPubMedWeb of Science
    1. Letsou A.,
    2. Arora K.,
    3. Wrana J. L.,
    4. Simin K.,
    5. Twombly V.,
    6. Jamal. J.,
    7. Staehling-Hampton K.,
    8. Hoffmann F. M.,
    9. Gelbart W. M.,
    10. Massague J.,
    11. O'Connor M. B.
    (1995) Drosophila Dpp signaling is mediated by the punt gene product: a dual ligand-binding type II receptor of the TGF beta receptor family. Cell 80, 899–908
    OpenUrlCrossRefPubMedWeb of Science
    1. Liu F.,
    2. Hata A.,
    3. Baker J. C.,
    4. Doody J.,
    5. Carcamo J.,
    6. Harland R. M.,
    7. Massague J.
    (1996) A human mad protein acting as a BMP-regulated transcriptional activator. Nature 381, 620–623
    OpenUrlCrossRefPubMed
    1. Macias-Silva M.,
    2. Abdollah S.,
    3. Hoodless P. A.,
    4. Pirone R.,
    5. Attisano L.,
    6. Wrana J. L.
    (1996) MADR2 is a substrate of the TGFreceptor and its phosphorylation is required for nuclear accumulation and signaling. Cell 87, 1215–1224
    OpenUrlCrossRefPubMedWeb of Science
    1. Massague J.
    (1996) TGFsignaling: receptors, transducers, and mad proteins. Cell 85, 947–950
    OpenUrlCrossRefPubMedWeb of Science
    1. Nellen D.,
    2. Burke R.,
    3. Struhl G.,
    4. Basler K.
    (1996) Direct and long-range action of a DPP morphogen gradient. Cell 85, 357–368
    OpenUrlCrossRefPubMedWeb of Science
    1. Newfeld S. J.,
    2. Chartoff E. H.,
    3. Graff J. M.,
    4. Lemton D. A.,
    5. Gelbert W. M.
    (1996) Mothers against dpp encodes a conserved cytoplasmic protein required in DPP/TGF--responsive cells. Development 122, 2099–2108
    OpenUrlAbstract
    1. Nishimatsu S.,
    2. Suzuki A.,
    3. Shoda A.,
    4. Murakami K.,
    5. Ueno N.
    (1992) Genes for bone morphogenetic proteins are differentially transcribed in early amphibian embryos. Bioch. Biophys. Res. Comm 186, 1487–1495
    OpenUrlCrossRefPubMedWeb of Science
    1. Onichtchouk D.,
    2. Gawantka V.,
    3. Dosch R.,
    4. Delius H.,
    5. Hirschfeld.,
    6. Blumenstock C.,
    7. Niehrs C.
    (1996) The Xvent-2 homeobox gene is part of the BMP-4 signallingpathway controling dorsovental patterning of Xenopus mesoderm. Development 122, 3045–3053
    OpenUrlAbstract
    1. Piccolo S.,
    2. Sasai Y.,
    3. Lu B.,
    4. De Robertis E. M.
    (1996) Dorsovental patterning in Xenopus: inhibition of ventral signals by direct binding of chordin to BMP-4. Cell 86, 589–598
    OpenUrlCrossRefPubMedWeb of Science
    1. Raftery L. A.,
    2. Twombly V.,
    3. Wharton K.,
    4. Gelbert W. M.
    (1995) Genetic screens to identify elements of the decapentaplegic signaling pathway in Drosophila. Genetics 139, 241–254
    OpenUrlAbstract/FREE Full Text
    1. Rosenzweig B. L.,
    2. Imamura T.,
    3. Okadome T.,
    4. Cox G. N.,
    5. Yamashita H.,
    6. ten Dijke P.,
    7. Helding C. H.,
    8. Miyazono K.
    (1995) Cloning and characterization of a human type II receptor for bone morphogenetic proteins. Proc. Natl. Acad. Sci. USA 92, 7632–7636
    OpenUrlAbstract/FREE Full Text
    1. Savage C.,
    2. Das P.,
    3. Finelli A.,
    4. Townsend S.,
    5. Sun C.,
    6. Baird S.,
    7. Padgett R.
    (1996) Caenorhabditis elegans gene sma-2 and sma-3 and sma-4 define a conserved family of transforming growth factor b pathway components. Proc. Natl. Acad. Sci. USA 93, 7790–9974
    OpenUrl
    1. Sekelsky J. J.,
    2. Newfeld S. J.,
    3. Raftery L. A.,
    4. Chartoff E. H.,
    5. Gelbert W. M.
    (1995) Genetic characterization and cloning of Mothers against dpp, a gene required for decapentaplegic function in Drosophila melanogaster. Genetics 139, 1347–1358
    OpenUrlAbstract/FREE Full Text
    1. Schmidt J. E.,
    2. Suzuki A.,
    3. Ueno N.,
    4. Kimelman D.
    (1995) Localized BMP-4 mediates dorsal/ventral patterning in the early Xenopus embryo. Dev. Biol 169, 37–50
    OpenUrlCrossRefPubMedWeb of Science
    1. Shi Y.,
    2. Hata A.,
    3. Lo R. S.,
    4. Massague,
    5. Pavletich N. P.
    (1997) A structural basis for mutational inactivation of the tumor suppressor Smad4. Nature 388, 87–93
    OpenUrlCrossRefPubMedWeb of Science
    1. Smith J. C.
    (1995) Mesoderm-inducing factors and mesodermal patterning. Curr. Opin. Cell Biol 7, 856–61
    OpenUrlCrossRefPubMedWeb of Science
    1. Smith W. C.,
    2. McKendry R.,
    3. Ribisi S.,
    4. Harland R. M.
    (1995) A nodal-related gene defines a physical and functional domain within the Spemann organizer. Cell 82, 37–46
    OpenUrlCrossRefPubMedWeb of Science
    1. Steinbeisser H.,
    2. Fainsod A.,
    3. Niehrs C.,
    4. Sasai Y.,
    5. De Robertis E. M.
    (1995) The role of gsc and BMP-4 in dorsal-ventral patterning of the marginal zone in Xenopus: a loss-of-function study using antisense RNA. EMBO J 14, 230–243
    OpenUrl
    1. Suzuki A.,
    2. Nishimatsu S.,
    3. Murakami K.,
    4. Ueno N.
    (1993) Differential expression of Xenopous BMPs in early embryos and tissues. Zool. Sci 10, 175–178
    OpenUrlPubMedWeb of Science
    1. Suzuki A.,
    2. Thies R. S.,
    3. Yamaji N.,
    4. Song J. J.,
    5. Wozney J. M.,
    6. Murakami K.,
    7. Ueno N.
    (1994) A truncated bone morphogenetic protein receptor affects dorsal-ventral patterning in the early Xenopus embryo. Proc. Natl. Acad. Sci. USA 91, 10255–10259
    OpenUrlAbstract/FREE Full Text
    1. ten Dijke P.,
    2. Yamashita H.,
    3. Sampath T. K.,
    4. Reddi A. H.,
    5. Estevez M.,
    6. Riddle D. L.,
    7. Ichijo H.,
    8. Heldin C. H.,
    9. Miyazono K.
    (1994) Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J. Biol. Chem 269, 16985–16988
    OpenUrlAbstract/FREE Full Text
    1. Thomsen G. H.
    (1996) Xenopus mothers against decapentaplegic is an embryonic ventralizing agent that acts downstream of the BMP-2/4 receptor. Development 122, 2359–2366
    OpenUrlAbstract
    1. Thomsen G.,
    2. Melton D. A.
    (1993) Processed Vg1 protein is an axial mesoderm inducer in Xenopus. Cell 74, 433–441
    OpenUrlCrossRefPubMedWeb of Science
    1. Thomsen G.,
    2. Woolf T.,
    3. Whitman M.,
    4. Sokol S.,
    5. Vaughan J.,
    6. Vale W.,
    7. Melton D. A.
    (1990) Activins are expressed early in Xenopus embryogenesis and can induce axial mesoderm and anterior structures. Cell 63, 485–493
    OpenUrlCrossRefPubMedWeb of Science
    1. Watabe T.,
    2. Kim S.,
    3. Candia A.,
    4. Rothbächer U.,
    5. Hashimoto C.,
    6. Inoue K.,
    7. Cho K. W. Y.
    (1995) Molecular mechanisms of Spemann's organizer formation: conserved growth factor synergy between Xenopus and mouse. Genes Dev 9, 3038–3050
    OpenUrlAbstract/FREE Full Text
    1. Wall N. A.,
    2. Hogan B. L.
    (1994) TGF-related genes in development. Curr. Opin. Genet. Dev 4, 517–522
    OpenUrlCrossRefPubMed
    1. Wieser R.,
    2. Wrana J. L.,
    3. Massague J.
    (1995) GS domain mutations that constitutively activate T beta R-I, the downstream signaling component in the TGF-eta receptor complex. EMBO J 14, 2199–2208
    OpenUrlPubMedWeb of Science
    1. Wrana J. L.,
    2. Attisano L.,
    3. Wieser R.,
    4. Ventrua F.,
    5. Massague J.
    (1994) Mechanism of activation of the TGF-beta receptor. Nature 370, 341–347
    OpenUrlCrossRefPubMedWeb of Science
    1. Wu R.-Y.,
    2. Zhang Y.,
    3. Feng X.-H.,
    4. Derynck R.
    (1997) Homomeric and heteromeric interactions are required for signaling activity and functional cooperativity of Smad-3 and-4. Mol. Cell. Biol 17, 2521–2528
    OpenUrlAbstract/FREE Full Text
    1. Yamashita H.,
    2. ten Dijke P.,
    3. Huylebroeck D.,
    4. Sampath T. K.,
    5. Andries M.,
    6. Smith J. C.,
    7. Heldin C. H.,
    8. Miyazono K.
    (1995) Osteogenic protein-1 binds to activin type II receptors and induces certain activin-like effects. J. Cell Biol 130, 217–26
    OpenUrlAbstract/FREE Full Text
    1. Yingling J. M.,
    2. Das P.,
    3. Savage C.,
    4. Zhang M.,
    5. Padgetts R.,
    6. Wang X. F.
    (1996) Mammalian Dwarfins are phosphorylated in response to transforming growth factor-and are implicated in control of cell growth. Proc. Natl. Acad. Sci. USA 93, 8940–8944
    OpenUrlAbstract/FREE Full Text
    1. Zhang Y.,
    2. Feng X.-H.,
    3. Wu R.-Y.,
    4. Derynk R.
    (1996) Receptor-associated Mad homologues synergize as effectors of the TGF-response. Nature 383, 168–172
    OpenUrlCrossRefPubMedWeb of Science
    1. Zhang Y.,
    2. Musci T.,
    3. Derynck R.
    (1997) The tumor suppressor Smad 4/DPC 4 as a central mediator of Smad function. Current Biol 7, 270–276
    OpenUrlCrossRefPubMedWeb of Science
    1. Zimmerman L. B.,
    2. De Jesus-Escobar J. M.,
    3. Harland R. M.
    (1996) The Spemann organizer signal noggin binds and inactivates bone morphogenetic protein 4. Cell 86, 599–606
    OpenUrlCrossRefPubMedWeb of Science
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JOURNAL ARTICLES
Cellular interpretation of multiple TGF-beta signals: intracellular antagonism between activin/BVg1 and BMP-2/4 signaling mediated by Smads
A.F. Candia, T. Watabe, S.H. Hawley, D. Onichtchouk, Y. Zhang, R. Derynck, C. Niehrs, K.W. Cho
Development 1997 124: 4467-4480;
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