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Condensin and Repo-Man–PP1 co-operate in the regulation of chromosome architecture during mitosis

Nature Cell Biology volume 8, pages 1133–1142 (2006)Cite this article

Abstract

The reversible condensation of chromosomes during cell division remains a classic problem in cell biology. Condensation requires the condensin complex1 in certain experimental systems2,3,4,5,6,7,8, but not in many others9,10,11,12,13,14,15. Anaphase chromosome segregation almost always fails in condensin-depleted cells, leading to the formation of prominent chromatin bridges and cytokinesis failure4,9,10,11,12,13,14,15,16,17. Here, live-cell analysis of chicken DT40 cells bearing a conditional knockout of condensin subunit SMC2 revealed that condensin-depleted chromosomes abruptly lose their compact architecture during anaphase and form massive chromatin bridges. The compact chromosome structure can be preserved and anaphase chromosome segregation rescued by preventing the targeting subunit Repo-Man from recruiting protein phosphatase 1 (PP1) to chromatin at anaphase onset. This study identifies an activity critical for mitotic chromosome structure that is inactivated by Repo-Man–PP1 during anaphase. This activity, provisionally termed 'regulator of chromosome architecture' (RCA), cooperates with condensin to preserve the characteristic chromosome architecture during mitosis.

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Figure 1: Phenotypes of condensin-depleted cells in anaphase.
Figure 2: Overexpression of GFP–cyclin B3 rescues the anaphase bridging phenotype in condensin-depleted cells.
Figure 3: Expression of GFP–Repo-ManRAXA mutant suppresses the anaphase bridges in SMC2OFF cells.
Figure 4: Repo-Man is the critical factor regulated by CDK–cyclin B3.
Figure 5: Co-operation between condensin and CDKs in the establishment and maintenance of mitotic chromosome condensation.

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Acknowledgements

We thank L. Meijer for the gift of a panel of Cdk inhibitors, including roscovitine; A. Belmont for the gift of the Lac O/Lac I–GFP plasmids; T. Fukagawa for the gift of the CENP-H–GFP targeting construct; E. Nigg for the gift of anti-chicken cyclin B2; R. Tsien for the gift of mCherry; F. Kasai and M. Ferguson-Smith for the chicken Z-chromosome paint; J. Boudeau and D. Alessi for help with the CDK1 assays; and R. Allshire, M. Heck and N. Cobbe for comments on the manuscript. This work is supported by The Wellcome Trust (P.V., A.L. and W.C.E.), The Caledonian Research Foundation (D.H.) and Cancer Research-UK (J.M.S and C.J.F). W.C.E. and A.I.L. are Principal Research Fellows of The Wellcome Trust.

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Authors and Affiliations

  1. Wellcome Trust Centre for Cell Biology, Institute of Cell Biology, University of Edinburgh, Swann Building, King's Buildings, Mayfield Road, Edinburgh, EH9 3JR, UK

    Paola Vagnarelli, Damien F. Hudson, Susana A. Ribeiro, Fan Lai & William C. Earnshaw

  2. IPATIMUP, R. Roberto Frias, s/n, Porto, 4050-465, Portugal

    Susana A. Ribeiro

  3. MSI/WTB Complex, University of Dundee, Dow Street, Dundee, DD1 5EH, UK

    Laura Trinkle-Mulcahy & Angus I. Lamond

  4. Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK

    Jennifer M. Spence & Christine J. Farr

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Contributions

P.V., D.H., L.T.-M., F.L. and J.M.S. performed the experiments, which were planned by P.V., C.J.F., A.I.L. and W.C.E. The manuscript was written by P.V. and W.C.E. with contributions from all other auhors.

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Correspondence to William C. Earnshaw.

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Vagnarelli, P., Hudson, D., Ribeiro, S. et al. Condensin and Repo-Man–PP1 co-operate in the regulation of chromosome architecture during mitosis. Nat Cell Biol 8, 1133–1142 (2006). https://doi.org/10.1038/ncb1475

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