Skip to main content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
EMBO J. 1999 Jun 15; 18(12): 3404–3418.
PMCID: PMC1171420
PMID: 10369680

RYBP, a new repressor protein that interacts with components of the mammalian Polycomb complex, and with the transcription factor YY1.

Abstract

The products of the Polycomb group (PcG) of genes are necessary for the maintenance of transcriptional repression of a number of important developmental genes, including the homeotic genes. A two-hybrid screen was used to search for putative new members of the PcG of genes in mammals. We have identified a new Zn finger protein, RYBP, which interacts directly with both Ring1 proteins (Ring1A and Ring1B) and with M33, two mutually interacting sets of proteins of the mammalian Polycomb complex. Ring1 binds RYBP and M33 through the same C-terminal domain, whereas the RYBP-M33 interaction takes place through an M33 domain not involved in Ring1 binding. RYBP also interacts directly with YY1, a transcription factor partially related to the product of the Drosophila pleiohomeotic gene. In addition, we show here that RYBP acts as a transcriptional repressor in transiently transfected cells. Finally, RYBP shows a dynamic expression pattern during embryogenesis which initially overlaps partially that of Ring1A in the central nervous system, and later becomes ubiquitous. Taken together, these data suggest that RYBP may play a relevant role in PcG function in mammals.

Full Text

The Full Text of this article is available as a PDF (571K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  • Adler PN, Martin EC, Charlton J, Jones K. Phenotypic consequences and genetic interactions of a null mutation in the Drosophila Posterior Sex Combs gene. Dev Genet. 1991;12(5):349–361. [PubMed] [Google Scholar]
  • Akasaka T, Kanno M, Balling R, Mieza MA, Taniguchi M, Koseki H. A role for mel-18, a Polycomb group-related vertebrate gene, during theanteroposterior specification of the axial skeleton. Development. 1996 May;122(5):1513–1522. [PubMed] [Google Scholar]
  • Alkema MJ, van der Lugt NM, Bobeldijk RC, Berns A, van Lohuizen M. Transformation of axial skeleton due to overexpression of bmi-1 in transgenic mice. Nature. 1995 Apr 20;374(6524):724–727. [PubMed] [Google Scholar]
  • Alkema MJ, Bronk M, Verhoeven E, Otte A, van 't Veer LJ, Berns A, van Lohuizen M. Identification of Bmi1-interacting proteins as constituents of a multimeric mammalian polycomb complex. Genes Dev. 1997 Jan 15;11(2):226–240. [PubMed] [Google Scholar]
  • Alkema MJ, Jacobs J, Voncken JW, Jenkins NA, Copeland NG, Satijn DP, Otte AP, Berns A, van Lohuizen M. MPc2, a new murine homolog of the Drosophila polycomb protein is a member of the mouse polycomb transcriptional repressor complex. J Mol Biol. 1997 Nov 14;273(5):993–1003. [PubMed] [Google Scholar]
  • Bienz M, Müller J. Transcriptional silencing of homeotic genes in Drosophila. Bioessays. 1995 Sep;17(9):775–784. [PubMed] [Google Scholar]
  • Bornemann D, Miller E, Simon J. The Drosophila Polycomb group gene Sex comb on midleg (Scm) encodes a zinc finger protein with similarity to polyhomeotic protein. Development. 1996 May;122(5):1621–1630. [PubMed] [Google Scholar]
  • Brown JL, Mucci D, Whiteley M, Dirksen ML, Kassis JA. The Drosophila Polycomb group gene pleiohomeotic encodes a DNA binding protein with homology to the transcription factor YY1. Mol Cell. 1998 Jun;1(7):1057–1064. [PubMed] [Google Scholar]
  • Buchenau P, Hodgson J, Strutt H, Arndt-Jovin DJ. The distribution of polycomb-group proteins during cell division and development in Drosophila embryos: impact on models for silencing. J Cell Biol. 1998 Apr 20;141(2):469–481. [PMC free article] [PubMed] [Google Scholar]
  • Bunker CA, Kingston RE. Transcriptional repression by Drosophila and mammalian Polycomb group proteins in transfected mammalian cells. Mol Cell Biol. 1994 Mar;14(3):1721–1732. [PMC free article] [PubMed] [Google Scholar]
  • Busturia A, Bienz M. Silencers in abdominal-B, a homeotic Drosophila gene. EMBO J. 1993 Apr;12(4):1415–1425. [PMC free article] [PubMed] [Google Scholar]
  • Chatton B, Bahr A, Acker J, Kedinger C. Eukaryotic GST fusion vector for the study of protein-protein associations in vivo: application to interaction of ATFa with Jun and Fos. Biotechniques. 1995 Jan;18(1):142–145. [PubMed] [Google Scholar]
  • Cheng NN, Sinclair DA, Campbell RB, Brock HW. Interactions of polyhomeotic with Polycomb group genes of Drosophila melanogaster. Genetics. 1994 Dec;138(4):1151–1162. [PMC free article] [PubMed] [Google Scholar]
  • Chiang A, O'Connor MB, Paro R, Simon J, Bender W. Discrete Polycomb-binding sites in each parasegmental domain of the bithorax complex. Development. 1995 Jun;121(6):1681–1689. [PubMed] [Google Scholar]
  • Cohen KJ, Hanna JS, Prescott JE, Dang CV. Transformation by the Bmi-1 oncoprotein correlates with its subnuclear localization but not its transcriptional suppression activity. Mol Cell Biol. 1996 Oct;16(10):5527–5535. [PMC free article] [PubMed] [Google Scholar]
  • Coré N, Bel S, Gaunt SJ, Aurrand-Lions M, Pearce J, Fisher A, Djabali M. Altered cellular proliferation and mesoderm patterning in Polycomb-M33-deficient mice. Development. 1997 Feb;124(3):721–729. [PubMed] [Google Scholar]
  • Coulson M, Robert S, Eyre HJ, Saint R. The identification and localization of a human gene with sequence similarity to Polycomblike of Drosophila melanogaster. Genomics. 1998 Mar 15;48(3):381–383. [PubMed] [Google Scholar]
  • DeCamillis M, Cheng NS, Pierre D, Brock HW. The polyhomeotic gene of Drosophila encodes a chromatin protein that shares polytene chromosome-binding sites with Polycomb. Genes Dev. 1992 Feb;6(2):223–232. [PubMed] [Google Scholar]
  • Denisenko ON, Bomsztyk K. The product of the murine homolog of the Drosophila extra sex combs gene displays transcriptional repressor activity. Mol Cell Biol. 1997 Aug;17(8):4707–4717. [PMC free article] [PubMed] [Google Scholar]
  • Duncan IM. Polycomblike: a gene that appears to be required for the normal expression of the bithorax and antennapedia gene complexes of Drosophila melanogaster. Genetics. 1982 Sep;102(1):49–70. [PMC free article] [PubMed] [Google Scholar]
  • Evan GI, Lewis GK, Ramsay G, Bishop JM. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. [PMC free article] [PubMed] [Google Scholar]
  • Gindhart JG, Jr, Kaufman TC. Identification of Polycomb and trithorax group responsive elements in the regulatory region of the Drosophila homeotic gene Sex combs reduced. Genetics. 1995 Feb;139(2):797–814. [PMC free article] [PubMed] [Google Scholar]
  • Goodrich J. Plant development: Medea's maternal instinct. Curr Biol. 1998 Jul 2;8(14):R480–R484. [PubMed] [Google Scholar]
  • Green S, Issemann I, Sheer E. A versatile in vivo and in vitro eukaryotic expression vector for protein engineering. Nucleic Acids Res. 1988 Jan 11;16(1):369–369. [PMC free article] [PubMed] [Google Scholar]
  • Grossniklaus U, Vielle-Calzada JP, Hoeppner MA, Gagliano WB. Maternal control of embryogenesis by MEDEA, a polycomb group gene in Arabidopsis. Science. 1998 Apr 17;280(5362):446–450. [PubMed] [Google Scholar]
  • Gunning P, Leavitt J, Muscat G, Ng SY, Kedes L. A human beta-actin expression vector system directs high-level accumulation of antisense transcripts. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4831–4835. [PMC free article] [PubMed] [Google Scholar]
  • Gunster MJ, Satijn DP, Hamer KM, den Blaauwen JL, de Bruijn D, Alkema MJ, van Lohuizen M, van Driel R, Otte AP. Identification and characterization of interactions between the vertebrate polycomb-group protein BMI1 and human homologs of polyhomeotic. Mol Cell Biol. 1997 Apr;17(4):2326–2335. [PMC free article] [PubMed] [Google Scholar]
  • Hashimoto N, Brock HW, Nomura M, Kyba M, Hodgson J, Fujita Y, Takihara Y, Shimada K, Higashinakagawa T. RAE28, BMI1, and M33 are members of heterogeneous multimeric mammalian Polycomb group complexes. Biochem Biophys Res Commun. 1998 Apr 17;245(2):356–365. [PubMed] [Google Scholar]
  • Hemenway CS, Halligan BW, Levy LS. The Bmi-1 oncoprotein interacts with dinG and MPh2: the role of RING finger domains. Oncogene. 1998 May 14;16(19):2541–2547. [PubMed] [Google Scholar]
  • Hobert O, Jallal B, Ullrich A. Interaction of Vav with ENX-1, a putative transcriptional regulator of homeobox gene expression. Mol Cell Biol. 1996 Jun;16(6):3066–3073. [PMC free article] [PubMed] [Google Scholar]
  • Hobert O, Sures I, Ciossek T, Fuchs M, Ullrich A. Isolation and developmental expression analysis of Enx-1, a novel mouse Polycomb group gene. Mech Dev. 1996 Apr;55(2):171–184. [PubMed] [Google Scholar]
  • Holdeman R, Nehrt S, Strome S. MES-2, a maternal protein essential for viability of the germline in Caenorhabditis elegans, is homologous to a Drosophila Polycomb group protein. Development. 1998 Jul;125(13):2457–2467. [PubMed] [Google Scholar]
  • Hollenberg SM, Sternglanz R, Cheng PF, Weintraub H. Identification of a new family of tissue-specific basic helix-loop-helix proteins with a two-hybrid system. Mol Cell Biol. 1995 Jul;15(7):3813–3822. [PMC free article] [PubMed] [Google Scholar]
  • Ishida A, Asano H, Hasegawa M, Koseki H, Ono T, Yoshida MC, Taniguchi M, Kanno M. Cloning and chromosome mapping of the human Mel-18 gene which encodes a DNA-binding protein with a new 'RING-finger' motif. Gene. 1993 Jul 30;129(2):249–255. [PubMed] [Google Scholar]
  • Kalenik JL, Chen D, Bradley ME, Chen SJ, Lee TC. Yeast two-hybrid cloning of a novel zinc finger protein that interacts with the multifunctional transcription factor YY1. Nucleic Acids Res. 1997 Feb 15;25(4):843–849. [PMC free article] [PubMed] [Google Scholar]
  • Kanno M, Hasegawa M, Ishida A, Isono K, Taniguchi M. mel-18, a Polycomb group-related mammalian gene, encodes a transcriptional negative regulator with tumor suppressive activity. EMBO J. 1995 Nov 15;14(22):5672–5678. [PMC free article] [PubMed] [Google Scholar]
  • Kehle J, Beuchle D, Treuheit S, Christen B, Kennison JA, Bienz M, Müller J. dMi-2, a hunchback-interacting protein that functions in polycomb repression. Science. 1998 Dec 4;282(5395):1897–1900. [PubMed] [Google Scholar]
  • Kennison JA. The Polycomb and trithorax group proteins of Drosophila: trans-regulators of homeotic gene function. Annu Rev Genet. 1995;29:289–303. [PubMed] [Google Scholar]
  • Korf I, Fan Y, Strome S. The Polycomb group in Caenorhabditis elegans and maternal control of germline development. Development. 1998 Jul;125(13):2469–2478. [PubMed] [Google Scholar]
  • Kyba M, Brock HW. The Drosophila polycomb group protein Psc contacts ph and Pc through specific conserved domains. Mol Cell Biol. 1998 May;18(5):2712–2720. [PMC free article] [PubMed] [Google Scholar]
  • Kyba M, Brock HW. The SAM domain of polyhomeotic, RAE28, and scm mediates specific interactions through conserved residues. Dev Genet. 1998;22(1):74–84. [PubMed] [Google Scholar]
  • Luckow B, Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. [PMC free article] [PubMed] [Google Scholar]
  • Martin EC, Adler PN. The Polycomb group gene Posterior Sex Combs encodes a chromosomal protein. Development. 1993 Feb;117(2):641–655. [PubMed] [Google Scholar]
  • Mihaly J, Mishra RK, Karch F. A conserved sequence motif in Polycomb-response elements. Mol Cell. 1998 Jun;1(7):1065–1066. [PubMed] [Google Scholar]
  • Niman HL, Houghten RA, Walker LE, Reisfeld RA, Wilson IA, Hogle JM, Lerner RA. Generation of protein-reactive antibodies by short peptides is an event of high frequency: implications for the structural basis of immune recognition. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4949–4953. [PMC free article] [PubMed] [Google Scholar]
  • Nomura M, Takihara Y, Shimada K. Isolation and characterization of retinoic acid-inducible cDNA clones in F9 cells: one of the early inducible clones encodes a novel protein sharing several highly homologous regions with a Drosophila polyhomeotic protein. Differentiation. 1994 Jun;57(1):39–50. [PubMed] [Google Scholar]
  • Orlando V, Jane EP, Chinwalla V, Harte PJ, Paro R. Binding of trithorax and Polycomb proteins to the bithorax complex: dynamic changes during early Drosophila embryogenesis. EMBO J. 1998 Sep 1;17(17):5141–5150. [PMC free article] [PubMed] [Google Scholar]
  • Ou SK, Hwang JM, Patterson PH. A modified method for obtaining large amounts of high titer polyclonal ascites fluid. J Immunol Methods. 1993 Sep 27;165(1):75–80. [PubMed] [Google Scholar]
  • Pearce JJ, Singh PB, Gaunt SJ. The mouse has a Polycomb-like chromobox gene. Development. 1992 Apr;114(4):921–929. [PubMed] [Google Scholar]
  • Pirrotta V. Chromatin-silencing mechanisms in Drosophila maintain patterns of gene expression. Trends Genet. 1997 Aug;13(8):314–318. [PubMed] [Google Scholar]
  • Pirrotta V. Polycombing the genome: PcG, trxG, and chromatin silencing. Cell. 1998 May 1;93(3):333–336. [PubMed] [Google Scholar]
  • Pirrotta V, Chan CS, McCabe D, Qian S. Distinct parasegmental and imaginal enhancers and the establishment of the expression pattern of the Ubx gene. Genetics. 1995 Dec;141(4):1439–1450. [PMC free article] [PubMed] [Google Scholar]
  • Rastelli L, Chan CS, Pirrotta V. Related chromosome binding sites for zeste, suppressors of zeste and Polycomb group proteins in Drosophila and their dependence on Enhancer of zeste function. EMBO J. 1993 Apr;12(4):1513–1522. [PMC free article] [PubMed] [Google Scholar]
  • Reijnen MJ, Hamer KM, den Blaauwen JL, Lambrechts C, Schoneveld I, van Driel R, Otte AP. Polycomb and bmi-1 homologs are expressed in overlapping patterns in Xenopus embryos and are able to interact with each other. Mech Dev. 1995 Sep;53(1):35–46. [PubMed] [Google Scholar]
  • Rupp RA, Snider L, Weintraub H. Xenopus embryos regulate the nuclear localization of XMyoD. Genes Dev. 1994 Jun 1;8(11):1311–1323. [PubMed] [Google Scholar]
  • Sadowski I, Ptashne M. A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Res. 1989 Sep 25;17(18):7539–7539. [PMC free article] [PubMed] [Google Scholar]
  • Satijn DP, Otte AP. RING1 interacts with multiple Polycomb-group proteins and displays tumorigenic activity. Mol Cell Biol. 1999 Jan;19(1):57–68. [PMC free article] [PubMed] [Google Scholar]
  • Satijn DP, Gunster MJ, van der Vlag J, Hamer KM, Schul W, Alkema MJ, Saurin AJ, Freemont PS, van Driel R, Otte AP. RING1 is associated with the polycomb group protein complex and acts as a transcriptional repressor. Mol Cell Biol. 1997 Jul;17(7):4105–4113. [PMC free article] [PubMed] [Google Scholar]
  • Satijn DP, Olson DJ, van der Vlag J, Hamer KM, Lambrechts C, Masselink H, Gunster MJ, Sewalt RG, van Driel R, Otte AP. Interference with the expression of a novel human polycomb protein, hPc2, results in cellular transformation and apoptosis. Mol Cell Biol. 1997 Oct;17(10):6076–6086. [PMC free article] [PubMed] [Google Scholar]
  • Saurin AJ, Shiels C, Williamson J, Satijn DP, Otte AP, Sheer D, Freemont PS. The human polycomb group complex associates with pericentromeric heterochromatin to form a novel nuclear domain. J Cell Biol. 1998 Aug 24;142(4):887–898. [PMC free article] [PubMed] [Google Scholar]
  • Schoorlemmer J, Marcos-Gutiérrez C, Were F, Martínez R, García E, Satijn DP, Otte AP, Vidal M. Ring1A is a transcriptional repressor that interacts with the Polycomb-M33 protein and is expressed at rhombomere boundaries in the mouse hindbrain. EMBO J. 1997 Oct 1;16(19):5930–5942. [PMC free article] [PubMed] [Google Scholar]
  • Sewalt RG, van der Vlag J, Gunster MJ, Hamer KM, den Blaauwen JL, Satijn DP, Hendrix T, van Driel R, Otte AP. Characterization of interactions between the mammalian polycomb-group proteins Enx1/EZH2 and EED suggests the existence of different mammalian polycomb-group protein complexes. Mol Cell Biol. 1998 Jun;18(6):3586–3595. [PMC free article] [PubMed] [Google Scholar]
  • Sewalt RG, Gunster MJ, van der Vlag J, Satijn DP, Otte AP. C-Terminal binding protein is a transcriptional repressor that interacts with a specific class of vertebrate Polycomb proteins. Mol Cell Biol. 1999 Jan;19(1):777–787. [PMC free article] [PubMed] [Google Scholar]
  • Shi Y, Seto E, Chang LS, Shenk T. Transcriptional repression by YY1, a human GLI-Krüppel-related protein, and relief of repression by adenovirus E1A protein. Cell. 1991 Oct 18;67(2):377–388. [PubMed] [Google Scholar]
  • Shumacher A, Faust C, Magnuson T. Positional cloning of a global regulator of anterior-posterior patterning in mice. Nature. 1996 Sep 19;383(6597):250–253. [PubMed] [Google Scholar]
  • Simon J. Locking in stable states of gene expression: transcriptional control during Drosophila development. Curr Opin Cell Biol. 1995 Jun;7(3):376–385. [PubMed] [Google Scholar]
  • Simon J, Chiang A, Bender W, Shimell MJ, O'Connor M. Elements of the Drosophila bithorax complex that mediate repression by Polycomb group products. Dev Biol. 1993 Jul;158(1):131–144. [PubMed] [Google Scholar]
  • Sinclair DA, Milne TA, Hodgson JW, Shellard J, Salinas CA, Kyba M, Randazzo F, Brock HW. The Additional sex combs gene of Drosophila encodes a chromatin protein that binds to shared and unique Polycomb group sites on polytene chromosomes. Development. 1998 Apr;125(7):1207–1216. [PubMed] [Google Scholar]
  • Soto MC, Chou TB, Bender W. Comparison of germline mosaics of genes in the Polycomb group of Drosophila melanogaster. Genetics. 1995 May;140(1):231–243. [PMC free article] [PubMed] [Google Scholar]
  • Spanopoulou E, Zaitseva F, Wang FH, Santagata S, Baltimore D, Panayotou G. The homeodomain region of Rag-1 reveals the parallel mechanisms of bacterial and V(D)J recombination. Cell. 1996 Oct 18;87(2):263–276. [PubMed] [Google Scholar]
  • Stankunas K, Berger J, Ruse C, Sinclair DA, Randazzo F, Brock HW. The enhancer of polycomb gene of Drosophila encodes a chromatin protein conserved in yeast and mammals. Development. 1998 Oct;125(20):4055–4066. [PubMed] [Google Scholar]
  • Struhl G, Akam M. Altered distributions of Ultrabithorax transcripts in extra sex combs mutant embryos of Drosophila. EMBO J. 1985 Dec 1;4(12):3259–3264. [PMC free article] [PubMed] [Google Scholar]
  • Strutt H, Paro R. The polycomb group protein complex of Drosophila melanogaster has different compositions at different target genes. Mol Cell Biol. 1997 Dec;17(12):6773–6783. [PMC free article] [PubMed] [Google Scholar]
  • Strutt H, Cavalli G, Paro R. Co-localization of Polycomb protein and GAGA factor on regulatory elements responsible for the maintenance of homeotic gene expression. EMBO J. 1997 Jun 16;16(12):3621–3632. [PMC free article] [PubMed] [Google Scholar]
  • Takihara Y, Tomotsune D, Shirai M, Katoh-Fukui Y, Nishii K, Motaleb MA, Nomura M, Tsuchiya R, Fujita Y, Shibata Y, et al. Targeted disruption of the mouse homologue of the Drosophila polyhomeotic gene leads to altered anteroposterior patterning and neural crest defects. Development. 1997 Oct;124(19):3673–3682. [PubMed] [Google Scholar]
  • Tetsu O, Ishihara H, Kanno R, Kamiyasu M, Inoue H, Tokuhisa T, Taniguchi M, Kanno M. mel-18 negatively regulates cell cycle progression upon B cell antigen receptor stimulation through a cascade leading to c-myc/cdc25. Immunity. 1998 Oct;9(4):439–448. [PubMed] [Google Scholar]
  • van de Vosse E, Walpole SM, Nicolaou A, van der Bent P, Cahn A, Vaudin M, Ross MT, Durham J, Pavitt R, Wilkinson J, et al. Characterization of SCML1, a new gene in Xp22, with homology to developmental polycomb genes. Genomics. 1998 Apr 1;49(1):96–102. [PubMed] [Google Scholar]
  • van der Lugt NM, Domen J, Linders K, van Roon M, Robanus-Maandag E, te Riele H, van der Valk M, Deschamps J, Sofroniew M, van Lohuizen M, et al. Posterior transformation, neurological abnormalities, and severe hematopoietic defects in mice with a targeted deletion of the bmi-1 proto-oncogene. Genes Dev. 1994 Apr 1;8(7):757–769. [PubMed] [Google Scholar]
  • van Lohuizen M, Frasch M, Wientjens E, Berns A. Sequence similarity between the mammalian bmi-1 proto-oncogene and the Drosophila regulatory genes Psc and Su(z)2. Nature. 1991 Sep 26;353(6342):353–355. [PubMed] [Google Scholar]
  • van Lohuizen M, Tijms M, Voncken JW, Schumacher A, Magnuson T, Wientjens E. Interaction of mouse polycomb-group (Pc-G) proteins Enx1 and Enx2 with Eed: indication for separate Pc-G complexes. Mol Cell Biol. 1998 Jun;18(6):3572–3579. [PMC free article] [PubMed] [Google Scholar]
  • Vojtek AB, Hollenberg SM, Cooper JA. Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell. 1993 Jul 16;74(1):205–214. [PubMed] [Google Scholar]
  • Wedeen C, Harding K, Levine M. Spatial regulation of Antennapedia and bithorax gene expression by the Polycomb locus in Drosophila. Cell. 1986 Mar 14;44(5):739–748. [PubMed] [Google Scholar]
  • Zink B, Paro R. In vivo binding pattern of a trans-regulator of homoeotic genes in Drosophila melanogaster. Nature. 1989 Feb 2;337(6206):468–471. [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

-