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The Drosophila melanogaster condensin subunit Cap-G interacts with the centromere-specific histone H3 variant CID

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Abstract

The centromere-specific histone H3 variant CENP-A plays a crucial role in kinetochore specification and assembly. We chose a genetic approach to identify interactors of the Drosophila CENP-A homolog CID. Overexpression of cid in the proliferating eye imaginal disk results in a rough eye phenotype, which is dependent on the ability of the overexpressed protein to localize to the kinetochore. A screen for modifiers of the rough eye phenotype identified mutations in the Drosophila condensin subunit gene Cap-G as interactors. Yeast two-hybrid experiments also reveal an interaction between CID and Cap-G. While chromosome condensation in Cap-G mutant embryos appears largely unaffected, massive defects in sister chromatid segregation occur during mitosis. Taken together, our results suggest a link between the chromatin condensation machinery and kinetochore structure.

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References

  • Ahmad K, Henikoff S (2002) Histone H3 variants specify modes of chromatin assembly. Proc Natl Acad Sci U S A 99 (Suppl 4):16477–16484

    Article  CAS  PubMed  Google Scholar 

  • Andrade MA, Perez-Iratxeta C, Ponting CP (2001) Protein repeats: structures, functions, and evolution. J Struct Biol 134:117–131

    Article  CAS  PubMed  Google Scholar 

  • Aono N, Sutani T, Tomonaga T, Mochida S, Yanagida M (2002) Cnd2 has dual roles in mitotic condensation and interphase. Nature 417:197–202

    Article  CAS  PubMed  Google Scholar 

  • Ball AR Jr, Schmiesing JA, Zhou C, Gregson HC, Okada Y, Doi T, Yokomori K (2002) Identification of a chromosome-targeting domain in the human condensin subunit CNAP1/hCAP-D2/Eg7. Mol Cell Biol 22:5769–5781

    Article  CAS  PubMed  Google Scholar 

  • 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 87:1103–1114

    Article  PubMed  Google Scholar 

  • Blower MD, Karpen GH (2001) The role of Drosophila CID in kinetochore formation, cell-cycle progression and heterochromatin interactions. Nat Cell Biol 3:730–739

    Article  CAS  PubMed  Google Scholar 

  • Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415

    CAS  PubMed  Google Scholar 

  • Chuang PT, Albertson DG, Meyer BJ (1994) DPY-27: a chromosome condensation protein homolog that regulates C. elegans dosage compensation through association with the X chromosome. Cell 79:459–474

    Article  CAS  PubMed  Google Scholar 

  • Cleveland DW, Mao Y, Sullivan KF (2003) Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling. Cell 112:407–421

    Article  CAS  PubMed  Google Scholar 

  • Coelho PA, Queiroz-Machado J, Sunkel CE (2003) Condensin-dependent localisation of topoisomerase II to an axial chromosomal structure is required for sister chromatid resolution during mitosis. J Cell Sci 116:4763–4776

    Article  CAS  PubMed  Google Scholar 

  • Draviam VM, Xie S, Sorger PK (2004) Chromosome segregation and genomic stability. Curr Opin Genet Dev 14:120–125

    Article  CAS  PubMed  Google Scholar 

  • Evan GI, Lewis GK, Ramsay G, Bishop JM (1985) Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol 5:3610–3616

    CAS  PubMed  Google Scholar 

  • Freeman M (1996) Reiterative use of the EGF receptor triggers differentiation of all cell types in the Drosophila eye. Cell 87:651–660

    Article  CAS  PubMed  Google Scholar 

  • Fukagawa T (2004) Assembly of kinetochores in vertebrate cells. Exp Cell Res 296:21–27

    Article  CAS  PubMed  Google Scholar 

  • Hagstrom KA, Holmes VF, Cozzarelli NR, Meyer BJ (2002) C. elegans condensin promotes mitotic chromosome architecture, centromere organization, and sister chromatid segregation during mitosis and meiosis. Genes Dev 16:729–742

    Article  CAS  PubMed  Google Scholar 

  • Hagstrom KA, Meyer BJ (2003) Condensin and cohesin: more than chromosome compactor and glue. Nat Rev Genet 4:520–534

    Article  CAS  PubMed  Google Scholar 

  • Hazelett DJ, Bourouis M, Walldorf U, Treisman JE (1998) decapentaplegic and wingless are regulated by eyes absent and eyegone and interact to direct the pattern of retinal differentiation in the eye disc. Development 125:3741–3751

    CAS  PubMed  Google Scholar 

  • Henikoff S, Ahmad K, Platero JS, van Steensel B (2000) Heterochromatic deposition of centromeric histone H3-like proteins. Proc Natl Acad Sci U S A 97:716–721

    CAS  PubMed  Google Scholar 

  • Hirano T (2000) Chromosome cohesion, condensation, and separation. Annu Rev Biochem 69:115–144

    Article  CAS  PubMed  Google Scholar 

  • Hirano T, Kobayashi R, Hirano M (1997) Condensins, chromosome condensation protein complexes containing XCAP-C, XCAP-E and a Xenopus homolog of the Drosophila Barren protein. Cell 89:511–521

    Article  CAS  PubMed  Google Scholar 

  • Horn C, Wimmer EA (2000) A versatile vector set for animal transgenesis. Dev Genes Evol 210:630–637

    Article  CAS  PubMed  Google Scholar 

  • Howman EV, Fowler KJ, Newson AJ, Redward S, MacDonald AC, Kalitsis P, Choo KH (2000) Early disruption of centromeric chromatin organization in centromere protein A (Cenpa) null mice. Proc Natl Acad Sci U S A 97:1148–1153

    Article  CAS  PubMed  Google Scholar 

  • Hudson DF, Vagnarelli P, Gassmann R, Earnshaw WC (2003) Condensin is required for nonhistone protein assembly and structural integrity of vertebrate mitotic chromosomes. Dev Cell 5:323–336

    Article  CAS  PubMed  Google Scholar 

  • Jacobs H, Richter D, Venkatesh T, Lehner C (2002) Completion of mitosis requires neither fzr/rap nor fzr2, a male germline-specific Drosophila Cdh1 homolog. Curr Biol 12:1435–1441

    Article  CAS  PubMed  Google Scholar 

  • Jäger H, Herzig A, Lehner CF, Heidmann S (2001) Drosophila Separase is required for sister chromatid separation and binds to PIM and THR. Genes Dev 15:2572–2584

    Article  PubMed  Google Scholar 

  • Jäger H, Herzig B, Herzig A, Sticht H, Lehner CF, Heidmann S (2004) Structure predictions and interaction studies indicate homology of separase N-terminal regulatory domains and Drosophila THR. Cell Cycle 3:182–188

    PubMed  Google Scholar 

  • Knoblich JA, Lehner CF (1993) Synergistic action of Drosophila cyclin A and cyclin B during the G2-M transition. EMBO J 12:65–74

    CAS  PubMed  Google Scholar 

  • Lasko PF, Pardue ML (1988) Studies of the genetic organization of the vestigial microregion of Drosophila melanogaster. Genetics 120:495–502

    CAS  PubMed  Google Scholar 

  • Leismann O, Herzig A, Heidmann S, Lehner CF (2000) Degradation of Drosophila PIM regulates sister chromatid separation during mitosis. Genes Dev 14:2192–2205

    Article  CAS  PubMed  Google Scholar 

  • 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 7:127–136

    Article  CAS  PubMed  Google Scholar 

  • McAinsh AD, Tytell JD, Sorger PK (2003) Structure, function, and regulation of budding yeast kinetochores. Annu Rev Cell Dev Biol 19:519–539

    Article  CAS  PubMed  Google Scholar 

  • Meluh PB, Koshland D (1995) Evidence that the MIF2 gene of Saccharomyces cerevisiae encodes a centromere protein with homology to the mammalian centromere protein CENP-C. Mol Biol Cell 6:793–807

    CAS  PubMed  Google Scholar 

  • Moore LL, Roth MB (2001) HCP-4, a CENP-C-like protein in Caenorhabditis elegans, is required for resolution of sister centromeres. J Cell Biol 153:1199–1208

    Article  CAS  PubMed  Google Scholar 

  • Neuwald AF, Hirano T (2000) HEAT repeats associated with condensins, cohesins, and other complexes involved in chromosome-related functions. Genome Res 10:1445–1452

    Article  CAS  PubMed  Google Scholar 

  • Nishihashi A, Haraguchi T, Hiraoka Y, Ikemura T, Regnier V, Dodson H, Earnshaw WC, Fukagawa T (2002) CENP-I is essential for centromere function in vertebrate cells. Dev Cell 2:463–476

    Article  CAS  PubMed  Google Scholar 

  • Oegema K, Desai A, Rybina S, Kirkham M, Hyman AA (2001) Functional analysis of kinetochore assembly in Caenorhabditis elegans. J Cell Biol 153:1209–1226

    Article  CAS  PubMed  Google Scholar 

  • Ono T, Losada A, Hirano M, Myers MP, Neuwald AF, Hirano T (2003) Differential contributions of condensin I and condensin II to mitotic chromosome architecture in vertebrate cells. Cell 115:109–121

    Article  CAS  PubMed  Google Scholar 

  • Rørth P (1998) Gal4 in the Drosophila female germline. Mech Dev 78:113–118

    Article  PubMed  Google Scholar 

  • Shelby RD, Vafa O, Sullivan KF (1997) Assembly of CENP-A into centromeric chromatin requires a cooperative array of nucleosomal DNA contact sites. J Cell Biol 136:501–513

    Article  CAS  PubMed  Google Scholar 

  • Stear JH, Roth MB (2002) Characterization of HCP-6, a C. elegans protein required to prevent chromosome twisting and merotelic attachment. Genes Dev 16:1498–1508

    Article  CAS  PubMed  Google Scholar 

  • Steffensen S, Coelho PA, Cobbe N, Vass S, Costa M, Hassan B, Prokopenko SN, Bellen H, Heck MM, Sunkel CE (2001) A role for Drosophila SMC4 in the resolution of sister chromatids in mitosis. Curr Biol 11:295–307

    Article  CAS  PubMed  Google Scholar 

  • Sullivan BA, Blower MD, Karpen GH (2001) Determining centromere identity: cyclical stories and forking paths. Nat Rev Genet 2:584–596

    Article  CAS  PubMed  Google Scholar 

  • Swedlow JR, Hirano T (2003) The making of the mitotic chromosome: modern insights into classical questions. Mol Cell 11:557–569

    Article  CAS  PubMed  Google Scholar 

  • Tseng AS, Hariharan IK (2002) An overexpression screen in Drosophila for genes that restrict growth or cell-cycle progression in the developing eye. Genetics 162:229–243

    CAS  PubMed  Google Scholar 

  • Van Hooser AA, Ouspenski, II, Gregson HC, Starr DA, Yen TJ, Goldberg ML, Yokomori K, Earnshaw WC, Sullivan KF, Brinkley BR (2001) Specification of kinetochore-forming chromatin by the histone H3 variant CENP-A. J Cell Sci 114:3529–3542

    CAS  PubMed  Google Scholar 

  • Weiss A, Herzig A, Jacobs H, Lehner CF (1998) Continuous Cyclin E expression inhibits progression through endoreduplication cycles in Drosophila. Curr Biol 8:239–242

    Article  CAS  PubMed  Google Scholar 

  • Westermann S, Cheeseman IM, Anderson S, Yates JR III, Drubin DG, Barnes G (2003) Architecture of the budding yeast kinetochore reveals a conserved molecular core. J Cell Biol 163:215–222

    Article  CAS  PubMed  Google Scholar 

  • Wignall SM, Deehan R, Maresca TJ, Heald R (2003) The condensin complex is required for proper spindle assembly and chromosome segregation in Xenopus egg extracts. J Cell Biol 161:1041–1051

    Article  CAS  PubMed  Google Scholar 

  • Wodarz A, Hinz U, Engelbert M, Knust E (1995) Expression of crumbs confers apical character on plasma-membrane domains of ectodermal epithelia of Drosophila. Cell 82:67–76

    Article  CAS  PubMed  Google Scholar 

  • Yeong FM, Hombauer H, Wendt KS, Hirota T, Mudrak I, Mechtler K, Loregger T, Marchler-Bauer A, Tanaka K, Peters JM, Ogris E (2003) Identification of a subunit of a novel Kleisin-beta/SMC complex as a potential substrate of protein phosphatase 2A. Curr Biol 13:2058–2064

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Katharina Neugebauer for technical support and C.-ting Wu, Steve DiNardo, and Volker Hartenstein for fly stocks. We are indebted to Thom Kaufman for providing the cid mutant lines prior to publication. We thank members of the laboratory for helpful discussions and Christian F. Lehner for generous support and critical reading of the manuscript. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (He 2354-1).

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  1. Hubert Jäger

    Present address: Astra-Zeneca Ltd, 22880, Wedel, Germany

  2. Melanie Rauch

    Present address: Department for Clinical and Biological Sciences and Department of Molecular Immunology Pharmacenter, University of Basel, 4056, Basel, Switzerland

Authors and Affiliations

  1. Lehrstuhl für Genetik, University of Bayreuth, 95440, Bayreuth, Germany

    Hubert Jäger, Melanie Rauch & Stefan Heidmann

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  1. Hubert Jäger
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Correspondence to Stefan Heidmann.

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Communicated by F. Uhlmann

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Jäger, H., Rauch, M. & Heidmann, S. The Drosophila melanogaster condensin subunit Cap-G interacts with the centromere-specific histone H3 variant CID. Chromosoma 113, 350–361 (2005). https://doi.org/10.1007/s00412-004-0322-4

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  • DOI: https://doi.org/10.1007/s00412-004-0322-4

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