Chromosoma

, Volume 112, Issue 5, pp 231–239 | Cite as

A proposed role for the Polycomb group protein dRING in meiotic sister-chromatid cohesion

  • Eric M. Balicky
  • Lynn Young
  • Terry L. Orr-Weaver
  • Sharon E. Bickel
Research Article

Abstract

ORD protein is required for accurate chromosome segregation during male and female meiosis in Drosophila melanogaster. Null ord mutations result in random segregation of sister chromatids during both meiotic divisions because cohesion is completely abolished prior to kinetochore capture of microtubules during meiosis I. Previous analyses of mutant ord alleles have led us to propose that the C-terminal half of the ORD protein mediates protein-protein interactions that are essential for sister-chromatid cohesion. To identify proteins that interact with ORD, we conducted a yeast two-hybrid screen using an ORD bait and isolated dRING, a core subunit of the Drosophila Polycomb repressive complex 1. We show that a missense mutation in ORD completely ablates the two-hybrid interaction with dRING and prevents nuclear retention of the mutant ORD protein in male meiotic cells. Using affinity-purified antibodies generated against full-length recombinant dRING, we demonstrate that dRING protein is expressed in the male and female gonads and colocalizes extensively with ORD on the chromatin of primary spermatocytes during G2 of meiosis. Our results suggest a novel role for the Polycomb group protein dRING and are consistent with the model that interaction of dRING and ORD is required to promote the proper segregation of meiotic chromosomes.

References

  1. Balicky EM, Endres MW, Lai C, Bickel SE (2002) Meiotic cohesion requires accumulation of ORD on chromosomes prior to condensation. Mol Biol Cell 21:3890–3900CrossRefGoogle Scholar
  2. Bhat MA, Philp AV, Glover DM, Bellen HJ (1996) Chromatid segregation at anaphase requires the barren product, a novel chromosome-associated protein that interacts with Topoisomerase II. Cell 87:1103–1114PubMedGoogle Scholar
  3. Bickel SE, Orr-Weaver TL (1998) Regulation of sister-chromatid cohesion during Drosophila meiosis. In: Zirkin BR (ed) Germ cell development, division, disruption and death. Springer, New York, pp 37–48Google Scholar
  4. Bickel SE, Wyman DW, Miyazaki WY, Moore DP, Orr-Weaver TL (1996) Identification of ORD, a Drosophila protein essential for sister-chromatid cohesion. EMBO J 15:1451–1459PubMedGoogle Scholar
  5. Bickel SE, Wyman DW, Orr-Weaver TL (1997) Mutational analysis of the Drosophila sister-chromatid cohesion protein ORD and its role in the maintenance of centromeric cohesion. Genetics 146:1319–1331PubMedGoogle Scholar
  6. Brent R, Ptashne M (1984) A bacterial repressor protein or a yeast transcriptional terminator can block upstream activation of a yeast gene. Nature 312:612–615PubMedGoogle Scholar
  7. Buonomo SB, Clyne RK, Fuchs J, Loidl J, Uhlmann F, Nasmyth K (2000) Disjunction of homologous chromosomes in meiosis I depends on proteolytic cleavage of the meiotic cohesin Rec8 by separin. Cell 103:387–398PubMedGoogle Scholar
  8. Cenci G, Bonaccorsi S, Pisano C, Verni F, Gatti M (1994) Chromatin and microtubule organization during premeiotic, meiotic, and early postmeiotic stages of Drosophila melanogaster spermatogenesis. J Cell Sci 107:3521–3534PubMedGoogle Scholar
  9. Dahiya A, Wong S, Gonzalo S, Gavin M, Dean DC (2001) Linking the Rb and polycomb pathways. Mol Cell 8:557–569PubMedGoogle Scholar
  10. Dietzel S, Niemann H, Bruckner B, Maurange C, Paro R (1999) The nuclear distribution of Polycomb during Drosophila melanogaster development shown with a GFP fusion protein. Chromosoma 108:83–94Google Scholar
  11. Fincham JRS (1966) Genetic complementation. Benjamin, New YorkGoogle Scholar
  12. Finley RL, Thomas BJ, Zipursky SL, Brent R (1996) Isolation of Drosophila cyclin D, a protein expressed in the morphogenetic furrow before entry into S phase. Proc Natl Acad Sci U S A 93:3011–3015Google Scholar
  13. Foster GG (1975) Negative complementation at the Notch locus of Drosophila melanogaster. Genetics 81:99–120PubMedGoogle Scholar
  14. Francis NJ, Kingston RE (2001) Mechanisms of transcriptional memory. Nat Rev Mol Cell Biol 2:409–421CrossRefPubMedGoogle Scholar
  15. Francis NJ, Saurin AJ, Shao Z, Kingston RE (2001) Reconstitution of a functional core Polycomb repressive complex. Mol Cell 8:545–556PubMedGoogle Scholar
  16. Fritsch C, Beuchle D, Muller J (2003) Molecular and genetic analysis of the Polycomb group gene Sex combs extra/Ring in Drosophila. Mech Dev 120:949–954CrossRefPubMedGoogle Scholar
  17. Goldstein LSB (1980) Mechanisms of chromosome orientation revealed by two meiotic mutants in Drosophila melanogaster. Chromosoma 78:79–111PubMedGoogle Scholar
  18. Golemis EA, Serebriiskii I, Gyuris J, Brent R (1997) Interaction trap/two-hybrid system to identify interacting proteins. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (ed) Current protocols in molecular biology. John Wiley and Sons, New York, pp 20.21.21–20.21.35Google Scholar
  19. Gyuris J, Golemis E, Chertkov H, Brent R (1993) Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell 75:791–803PubMedGoogle Scholar
  20. Hakimi MA, Bochar DA, Schmiesing JA, Dong Y, Barak OG, Speicher DW, Yokomori K, Shiekhattar R (2002) A chromatin remodelling complex that loads cohesin onto human chromosomes. Nature 418:994–998CrossRefPubMedGoogle Scholar
  21. Hawley RS, Walker MY (2003) Advanced genetic analysis: finding meaning in a genome. Blackwell Science, Malden, MassGoogle Scholar
  22. Jacobs JJ, van Lohuizen M (2002) Polycomb repression: from cellular memory to cellular proliferation and cancer. Biochim Biophys Acta 1602:151–161CrossRefPubMedGoogle Scholar
  23. Kudo N, Matsumori N, Taoka H, Fujiwara D, Schreiner E, Wolff B, Yoshida M, Horinouchi S (1999) Leptomycin B inactivates CRM1/exportin 1 by covalent modification at a cysteine residue in the central conserved region. Proc Natl Acad Sci U S A 96:9112–9117Google Scholar
  24. Kunkel TA (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A 82:488–492Google Scholar
  25. Lee JY, Orr-Weaver TL (2001) The molecular basis of sister-chromatid cohesion. Annu Rev Cell Dev Biol 17:753–777PubMedGoogle Scholar
  26. Levine SS, Weiss A, Erdjument-Bromage H, Shao Z, Tempst P, Kingston RE (2002) The core of the Polycomb repressive complex is compositionally and functionally conserved in flies and humans. Mol Cell Biol 22:6070–6078CrossRefPubMedGoogle Scholar
  27. Losada A, Hirano T (2001) Shaping the metaphase chromosome: coordination of cohesion and condensation. Bioessays 23:924–935CrossRefPubMedGoogle Scholar
  28. Lupo R, Breiling A, Bianchi ME, Orlando V (2001) Drosophila chromosome condensation proteins Topoisomerase II and Barren colocalize with Polycomb and maintain Fab-7 PRE silencing. Mol Cell 7:127–136PubMedGoogle Scholar
  29. Mason JM (1976) Orientation disruptor ( ord): a recombination-defective and disjunction-defective meiotic mutant in Drosophila melanogaster. Genetics 84:545–572PubMedGoogle Scholar
  30. Mihaly J, Mishra RK, Karch F (1998) A conserved sequence motif in Polycomb-response elements. Mol Cell 1:1065–1066PubMedGoogle Scholar
  31. Miyagishima H, Isono K, Fujimura Y, Iyo M, Takihara Y, Masumoto H, Vidal M, Koseki H (2003) Dissociation of mammalian Polycomb-group proteins, Ring1B and Rae28/Ph1, from the chromatin correlates with configuration changes of the chromatin in mitotic and meiotic prophase. Histochem Cell Biol 120:111–119CrossRefPubMedGoogle Scholar
  32. Miyazaki WY, Orr-Weaver TL (1992) Sister-chromatid misbehavior in Drosophila ord mutants. Genetics 132:1047–1061PubMedGoogle Scholar
  33. Orlando V (2003) Polycomb, epigenomes, and control of cell identity. Cell 112:599–606PubMedGoogle Scholar
  34. Petronczki M, Siomos MF, Nasmyth K (2003) Un menage a quatre: the molecular biology of chromosome segregation in meiosis. Cell 112:423–440PubMedGoogle Scholar
  35. Portin P (1975) Allelic negative complementation at the Abruptex locus of Drosophila melanogaster. Genetics 81:121–133PubMedGoogle Scholar
  36. Ringrose L, Paro R (2001) Remembering silence. Bioessays 23:566–570PubMedGoogle Scholar
  37. Satijn DP, Otte AP (1999) RING1 interacts with multiple Polycomb-group proteins and displays tumorigenic activity. Mol Cell Biol 19:57–68PubMedGoogle Scholar
  38. Satijn DP, Gunster MJ, van der Vlag J, Hamer KM, Schul W, Alkema MJ, Saurin AJ, Freemont PS, van Driel R, Otte AP (1997) RING1 is associated with the Polycomb group protein complex and acts as a transcriptional repressor. Mol Cell Biol 17:4105–4113PubMedGoogle Scholar
  39. Saurin AJ, Shao Z, Erdjument-Bromage H, Tempst P, Kingston RE (2001) A Drosophila Polycomb group complex includes Zeste and dTAFII proteins. Nature 412:655–660CrossRefPubMedGoogle Scholar
  40. Schoorlemmer J, Marcos-Gutierrez C, Were F, Martinez R, Garcia E, Satijn DP, Otte AP, Vidal M (1997) Ring1A is a transcriptional repressor that interacts with the Polycomb-M33 protein and is expressed at rhombomere boundaries in the mouse hindbrain. EMBO J 16:5930–5942CrossRefPubMedGoogle Scholar
  41. Shao Z, Raible F, Mollaaghababa R, Guyon JR, Wu CT, Bender W, Kingston RE (1999) Stabilization of chromatin structure by PRC1, a Polycomb complex. Cell 98:37–46PubMedGoogle Scholar
  42. Simon JA (2003) Polycomb group proteins. Curr Biol 13: R79–80CrossRefPubMedGoogle Scholar
  43. Siomos MF, Badrinath A, Pasierbek P, Livingstone D, White J, Glotzer M, Nasmyth K (2001) Separase is required for chromosome segregation during meiosis I in Caenorhabditis elegans. Curr Biol 11:1825–1835CrossRefPubMedGoogle Scholar
  44. Soto MC, Chou TB, Bender W (1995) Comparison of germline mosaics of genes in the Polycomb group of Drosophila melanogaster. Genetics 140:231–243PubMedGoogle Scholar
  45. Swedlow JR, Hirano T (2003) The making of the mitotic chromosome: modern insights into classical questions. Mol Cell 11:557–569PubMedGoogle Scholar
  46. Tuckfield A, Clouston DR, Wilanowski TM, Zhao LL, Cunningham JM, Jane SM (2002) Binding of the RING Polycomb proteins to specific target genes in complex with the grainyhead-like family of developmental transcription factors. Mol Cell Biol 22:1936–1946CrossRefPubMedGoogle Scholar
  47. Uhlmann F (2003) Chromosome cohesion and separation: from men and molecules. Curr Biol 13: R104–114CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Eric M. Balicky
    • 1
  • Lynn Young
    • 2
  • Terry L. Orr-Weaver
    • 2
  • Sharon E. Bickel
    • 1
  1. 1.Department of Biological SciencesDartmouth CollegeHanoverUSA
  2. 2.Whitehead Institute for Biomedical Research and Department of BiologyMassachusetts Institute of TechnologyCambridgeUSA

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