Physical and Functional Interaction of SMADs and p300/CBP*

SMADs are transforming growth factor β (TGF-β) receptor substrates and mediators of TGF-β transcriptional responses. Here we provide evidence that the coactivators p300 and CBP interact with Smads 1 through 4. The biological relevance of this interaction is shown in vivo by overexpression of the adenovirus E1A protein and mutant forms of E1A that lack p300-binding sites. Wild-type E1A, but not the mutants, inhibits SMAD-dependent transcriptional responses to TGF-β. E1A also inhibits the intrinsic transactivating function of the Smad4 MH2 domain. In addition, overexpression of p300 enhances SMAD-dependent transactivation. Our results suggest a role for p300/CBP in SMAD-mediated transcriptional activation and provide an explanation for the observed ability of E1A to interfere with TGF-β action.

[1]  J. Sekelsky,et al.  Genetic characterization and cloning of mothers against dpp, a gene required for decapentaplegic function in Drosophila melanogaster. , 1995, Genetics.

[2]  C. Mummery,et al.  Adenovirus E1A antagonizes both negative and positive growth signals elicited by transforming growth factor beta 1. , 1995, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[3]  E. Moran,et al.  Transforming growth factor beta 1 (TGF beta 1) reduces cellular levels of p34cdc2, and this effect is abrogated by adenovirus independently of the E1A-associated pRB binding activity. , 1992, Molecular biology of the cell.

[4]  P. D. Vize,et al.  Identification of a potential regulator of early transcriptional responses to mesoderm inducers in the frog embryo. , 1995, The EMBO journal.

[5]  J. Baker,et al.  A novel mesoderm inducer, Madr2, functions in the activin signal transduction pathway. , 1996, Genes & development.

[6]  Stephen H. Friend,et al.  Association between an oncogene and an anti-oncogene: the adenovirus E1A proteins bind to the retinoblastoma gene product , 1988, Nature.

[7]  P. Hoodless,et al.  MADR2 Is a Substrate of the TGFβ Receptor and Its Phosphorylation Is Required for Nuclear Accumulation and Signaling , 1996, Cell.

[8]  B. Howard,et al.  The Transcriptional Coactivators p300 and CBP Are Histone Acetyltransferases , 1996, Cell.

[9]  C. Heldin,et al.  Identification of Smad7, a TGFβ-inducible antagonist of TGF-β signalling , 1997, Nature.

[10]  C. Missero,et al.  Induction of transforming growth factor beta 1 resistance by the E1A oncogene requires binding to a specific set of cellular proteins. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Yigong Shi,et al.  The L3 loop: a structural motif determining specific interactions between SMAD proteins and TGF‐β receptors , 1998, The EMBO journal.

[12]  J. Massagué,et al.  Dual role of the Smad4/DPC4 tumor suppressor in TGFbeta-inducible transcriptional complexes. , 1997, Genes & development.

[13]  J. Massagué,et al.  Determinants of specificity in TGF-beta signal transduction. , 1998, Genes & development.

[14]  Xiao-Fan Wang,et al.  Tumor suppressor Smad4 is a transforming growth factor beta-inducible DNA binding protein , 1997, Molecular and cellular biology.

[15]  J. Graff,et al.  Xenopus Mad Proteins Transduce Distinct Subsets of Signals for the TGFβ Superfamily , 1996, Cell.

[16]  P. Hoodless,et al.  MADR1, a MAD-Related Protein That Functions in BMP2 Signaling Pathways , 1996, Cell.

[17]  R. Derynck,et al.  Receptor-associated Mad homologues synergize as effectors of the TGF-β response , 1996, Nature.

[18]  Xin Chen,et al.  A transcriptional partner for MAD proteins in TGF-β signalling , 1996, Nature.

[19]  Xiao-Fan Wang,et al.  Mammalian dwarfins are phosphorylated in response to transforming growth factor beta and are implicated in control of cell growth. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Kirby D. Johnson,et al.  Drosophila Mad binds to DNA and directly mediates activation of vestigial by Decapentaplegic , 1997, Nature.

[21]  Andrew J. Bannister,et al.  CBP‐induced stimulation of c‐Fos activity is abrogated by E1A. , 1995, The EMBO journal.

[22]  E. Harlow,et al.  Cellular targets for transformation by the adenovirus E1A proteins , 1989, Cell.

[23]  M. Kretzschmar,et al.  Opposing BMP and EGF signalling pathways converge on the TGF-β family mediator Smad1 , 1997, Nature.

[24]  Michael R. Green,et al.  Transcription activation by the adenovirus E1a protein , 1989, Nature.

[25]  L. Coussens,et al.  Transforming growth factor β1‐mediated induction of junB is selectively inhibited by expression of Ad.12‐E1A , 1994 .

[26]  Kohei Miyazono,et al.  TGF-β signalling from cell membrane to nucleus through SMAD proteins , 1997, Nature.

[27]  T. Tabata,et al.  Daughters against dpp modulates dpp organizing activity in Drosophila wing development , 1997, Nature.

[28]  J. Massagué,et al.  Mutations increasing autoinhibition inactivate tumour suppressors Smad2 and Smad4 , 1997, Nature.

[29]  P. Branton,et al.  Mapping of cellular protein-binding sites on the products of early-region 1A of human adenovirus type 5 , 1988, Molecular and cellular biology.

[30]  Ralf Janknecht,et al.  Transcriptional control: Versatile molecular glue , 1996, Current Biology.

[31]  J. Massagué,et al.  A human Mad protein acting as a BMP-regulated transcriptional activator , 1996, Nature.

[32]  J. Massagué,et al.  The TGF-beta family mediator Smad1 is phosphorylated directly and activated functionally by the BMP receptor kinase. , 1997, Genes & development.

[33]  J. Massagué.  TGF-beta signal transduction. , 1998, Annual review of biochemistry.

[34]  Yigong Shi,et al.  A structural basis for mutational inactivation of the tumour suppressor Smad4 , 1997, Nature.

[35]  P. Yaciuk,et al.  TGF-β1 inhibition of c-myc transcription and growth in keratinocytes is abrogated by viral transforming proteins with pRB binding domains , 1990, Cell.

[36]  Minoru Watanabe,et al.  Smad4 and FAST-1 in the assembly of activin-responsive factor , 1997, Nature.

[37]  E. Ziff,et al.  Complementary functions of E1a conserved region 1 cooperate with conserved region 3 to activate adenovirus serotype 5 early promoters , 1994, Journal of virology.

[38]  K. Miyazono,et al.  Smad6 inhibits signalling by the TGF-β superfamily , 1997, Nature.

[39]  J B Lawrence,et al.  Molecular cloning and functional analysis of the adenovirus E1A-associated 300-kD protein (p300) reveals a protein with properties of a transcriptional adaptor. , 1994, Genes & development.

[40]  C. Heldin,et al.  Identification of Smad2, a Human Mad-related Protein in the Transforming Growth Factor β Signaling Pathway* , 1997, The Journal of Biological Chemistry.

[41]  P. P. Hu,et al.  The viral oncoprotein E1A blocks transforming growth factor beta-mediated induction of p21/WAF1/Cip1 and p15/INK4B , 1997, Molecular and cellular biology.

[42]  N. Shiama.  The p300/CBP family: integrating signals with transcription factors and chromatin. , 1997, Trends in cell biology.

[43]  J. Massagué,et al.  Partnership between DPC4 and SMAD proteins in TGF-β signalling pathways , 1996, Nature.

[44]  J. Massagué,et al.  Smad6 inhibits BMP/Smad1 signaling by specifically competing with the Smad4 tumor suppressor. , 1998, Genes & development.

[45]  J. Wrana,et al.  The MAD-Related Protein Smad7 Associates with the TGFβ Receptor and Functions as an Antagonist of TGFβ Signaling , 1997, Cell.

[46]  R. Eckner.  p300 and CBP as transcriptional regulators and targets of oncogenic events. , 1996, Biological chemistry.

[47]  Jay H. Chang,et al.  Mechanism of E1A-Induced Transforming Growth Factor-β (TGF-β) Resistance in Mouse Keratinocytes Involves Repression of TGF-β Type II Receptor Transcription* , 1997, The Journal of Biological Chemistry.

[48]  Andrew J. Bannister,et al.  The CBP co-activator is a histone acetyltransferase , 1996, Nature.