Mad-max transcriptional repression is mediated by ternary complex formation with mammalian homologs of yeast repressor Sin3

The bHLH-ZIP protein Mad heterodimerizes with Max as a sequence-specific transcriptional repressor. Mad is rapidly induced upon differentiation, and the associated switch from Myc-Max to Mad-Max heterocomplexes seem to repress genes normally activated by Myc-Max. We have identified two related mammalian cDNAs that encode Mad-binding proteins. Both possess sequence homology with the yeast transcription repressor Sin3, including four conserved paired amphipathic helix (PAH) domains. mSin3A and mSin3B bind specifically to Mad and the related protein Mxi1. Mad-Max and mSin3 form ternary complexes in solution that specifically recognize the Mad-Max E box-binding site. Mad-mSin3 association requires PAH2 of mSin3A/mSin3B and the first 25 residues of Mad, which contains a putative amphipathic alpha-helical region. Point mutations in this region eliminate interaction with mSin3 proteins and block Mad transcriptional repression. We suggest that Mad-Max represses transcription by tethering mSin3 to DNA as corepressors and that a transcriptional repression mechanism is conserved from yeast to mammals.

[1]  I. Herskowitz,et al.  Activation of the yeast HO gene by release from multiple negative controls , 1987, Cell.

[2]  R. Roeder,et al.  Unliganded thyroid hormone receptor inhibits formation of a functional preinitiation complex: implications for active repression. , 1993, Genes & development.

[3]  F. Winston,et al.  Yeast SNF/SWI transcriptional activators and the SPT/SIN chromatin connection. , 1992, Trends in genetics : TIG.

[4]  Jonathan D. Licht,et al.  Drosophila Krüppel protein is a transcriptional represser , 1990, Nature.

[5]  A. Mitchell,et al.  Bipartite structure of an early meiotic upstream activation sequence from Saccharomyces cerevisiae , 1993, Molecular and cellular biology.

[6]  R. Eisenman,et al.  Myc and Max function as a nucleoprotein complex , 1992, Current Biology.

[7]  A. Duncan,et al.  The adenovirus E1A-associated 130-kD protein is encoded by a member of the retinoblastoma gene family and physically interacts with cyclins A and E. , 1993, Genes & development.

[8]  R. DePinho,et al.  Myc family oncoproteins function through a common pathway to transform normal cells in culture: cross-interference by Max and trans-acting dominant mutants. , 1992, Genes & development.

[9]  R. Eisenman,et al.  Transcriptional activities of the Myc and Max proteins in mammalian cells. , 1992, Current topics in microbiology and immunology.

[10]  R. Weinberg.  Tumor suppressor genes , 1993, Neuron.

[11]  M. Levine,et al.  Transcriptional repression of eukaryotic promoters , 1989, Cell.

[12]  R. Eisenman,et al.  Myc and Max proteins possess distinct transcriptional activities , 1992, Nature.

[13]  H. Wang,et al.  In vitro regulation of a SIN3-dependent DNA-binding activity by stimulatory and inhibitory factors. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[14]  S. Roth,et al.  The global transcriptional regulators, SSN6 and TUP1, play distinct roles in the establishment of a repressive chromatin structure. , 1994, Genes & development.

[15]  H. Wang,et al.  Transcriptional repression in Saccharomyces cerevisiae by a SIN3-LexA fusion protein , 1993, Molecular and cellular biology.

[16]  R. Eisenman,et al.  New light on Myc and Myb. Part I. Myc. , 1990, Genes & development.

[17]  M. Ewen,et al.  Molecular cloning, chromosomal mapping, and expression of the cDNA for p107, a retinoblastoma gene product-related protein , 1991, Cell.

[18]  W. Gu,et al.  Opposite regulation of gene transcription and cell proliferation by c-Myc and Max. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[19]  F. Hilger,et al.  Direct selection for mutants with increased K+ transport in Saccharomyces cerevisiae. , 1990, Genetics.

[20]  T. Alber.  Structure of the leucine zipper , 1992, Current Biology.

[21]  R. E. Esposito,et al.  RPD1 (SIN3/UME4) is required for maximal activation and repression of diverse yeast genes , 1991, Molecular and cellular biology.

[22]  R. DePinho,et al.  Suppression of Myc, but not E1a, transformation activity by Max-associated proteins, Mad and Mxi1. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Jonathan A. Cooper,et al.  Mammalian Ras interacts directly with the serine/threonine kinase raf , 1993, Cell.

[24]  M. Carlson,et al.  The N-Terminal TPR Region Is the Functional Domain of SSN6, a Nuclear Phosphoprotein of Saccharomyces cerevisiae , 1990, Molecular and cellular biology.

[25]  R. W. Davis,et al.  Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[26]  G. Evan,et al.  The c‐Myc protein induces cell cycle progression and apoptosis through dimerization with Max. , 1993, The EMBO journal.

[27]  M. Kozak.  The scanning model for translation: an update , 1989, The Journal of cell biology.

[28]  K. Struhl,et al.  Functional dissection of the yeast Cyc8–Tupl transcriptional co-repressor complex , 1994, Nature.

[29]  P R Nicholson,et al.  The Saccharomyces cerevisiae SIN3 gene, a negative regulator of HO, contains four paired amphipathic helix motifs , 1990, Molecular and cellular biology.

[30]  M. Eilers,et al.  The functions of Myc proteins. , 1992, Biochimica et biophysica acta.

[31]  L. Larsson,et al.  Expression of mad, mxi1, max and c-myc during induced differentiation of hematopoietic cells: opposite regulation of mad and c-myc. , 1994, Oncogene.

[32]  N. Hay,et al.  Sequence-specific transcriptional activation by Myc and repression by Max , 1993, Molecular and cellular biology.

[33]  S. Ho,et al.  Site-directed mutagenesis by overlap extension using the polymerase chain reaction. , 1989, Gene.

[34]  R. E. Esposito,et al.  Identification of negative regulatory genes that govern the expression of early meiotic genes in yeast. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Bruno Amati,et al.  Oncogenic activity of the c-Myc protein requires dimerization with Max , 1993, Cell.

[36]  D. Reinberg,et al.  DNA topoisomerase I is involved in both repression and activation of transcription , 1993, Nature.

[37]  R. Brent,et al.  Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. , 1993, Cell.

[38]  S. Fields,et al.  The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[39]  S. Madden,et al.  Transcriptional repression mediated by the WT1 Wilms tumor gene product. , 1991, Science.

[40]  G. Evan,et al.  Transcriptional activation by the human c-Myc oncoprotein in yeast requires interaction with Max , 1992, Nature.

[41]  R. Eisenman,et al.  Mad: A heterodimeric partner for Max that antagonizes Myc transcriptional activity , 1993, Cell.

[42]  S. Henikoff.  Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. , 1984, Gene.

[43]  D. Stillman,et al.  Both positive and negative regulators of HO transcription are required for mother-cell-specific mating-type switching in yeast , 1987, Cell.

[44]  J. Lopes,et al.  A pleiotropic phospholipid biosynthetic regulatory mutation in Saccharomyces cerevisiae is allelic to sin3 (sdi1, ume4, rpd1). , 1994, Genetics.

[45]  Roger Brent,et al.  Groucho is required for Drosophila neurogenesis, segmentation, and sex determination and interacts directly with hairy-related bHLH proteins , 1994, Cell.

[46]  R. Eisenman,et al.  A switch from Myc:Max to Mad:Max heterocomplexes accompanies monocyte/macrophage differentiation. , 1993, Genes & development.

[47]  J. Manley,et al.  Transcriptional repression by the Drosophila even-skipped protein: definition of a minimal repression domain. , 1993, Genes & development.

[48]  M. Levine,et al.  Short-range repression permits multiple enhancers to function autonomously within a complex promoter. , 1994, Genes & development.

[49]  Alexander D. Johnson,et al.  Ssn6-Tup1 is a general repressor of transcription in yeast , 1992, Cell.

[50]  J. Thorner,et al.  Mot1, a global repressor of RNA polymerase II transcription, inhibits TBP binding to DNA by an ATP-dependent mechanism. , 1994, Genes & development.

[51]  G D Schuler,et al.  A workbench for multiple alignment construction and analysis , 1991, Proteins.

[52]  A. Patel,et al.  myc function and regulation. , 1992, Annual review of biochemistry.

[53]  Adenovirus E1B oncoprotein tethers a transcriptional repression domain to p53. , 1994, Genes & development.

[54]  R. Renkawitz,et al.  A transferable silencing domain is present in the thyroid hormone receptor, in the v‐erbA oncogene product and in the retinoic acid receptor. , 1992, The EMBO journal.

[55]  S. Henikoff,et al.  Conserved arrangement of nested genes at the Drosophila Gart locus. , 1987, Genetics.