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Shugoshin collaborates with protein phosphatase 2A to protect cohesin

Nature volume 441, pages 46–52 (2006)Cite this article

Abstract

Sister chromatid cohesion, mediated by a complex called cohesin, is crucial—particularly at centromeres—for proper chromosome segregation in mitosis and meiosis. In animal mitotic cells, phosphorylation of cohesin promotes its dissociation from chromosomes, but centromeric cohesin is protected by shugoshin until kinetochores are properly captured by the spindle microtubules. However, the mechanism of shugoshin-dependent protection of cohesin is unknown. Here we find a specific subtype of serine/threonine protein phosphatase 2A (PP2A) associating with human shugoshin. PP2A colocalizes with shugoshin at centromeres and is required for centromeric protection. Purified shugoshin complex has an ability to reverse the phosphorylation of cohesin in vitro, suggesting that dephosphorylation of cohesin is the mechanism of protection at centromeres. Meiotic shugoshin of fission yeast also associates with PP2A, with both proteins collaboratively protecting Rec8-containing cohesin at centromeres. Thus, we have revealed a conserved mechanism of centromeric protection of eukaryotic chromosomes in mitosis and meiosis.

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Figure 1: PP2A associates and colocalizes with hSgo1.
Figure 2: PP2A is required for centromeric protection.
Figure 3: Interdependency of shugoshin and PP2A for localization.
Figure 4: Dephosphorylation of cohesin subunit SA.
Figure 5: S. pombe PP2A associating with Sgo1 is required for centromeric protection of Rec8-containing cohesin during meiosis I.
Figure 6: Individual ability of PP2A and shugoshin for centromeric protection.

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References

  1. Nasmyth, K. Disseminating the genome: joining, resolving, and separating sister chromatids during mitosis and meiosis. Annu. Rev. Genet. 35, 673–745 (2001)

    Article  CAS  Google Scholar 

  2. Milutinovich, M. & Koshland, D. E. SMC complexes–wrapped up in controversy. Science 300, 1101–1102 (2003)

    Article  CAS  Google Scholar 

  3. Hirano, T. Dynamic molecular linkers of the genome: the first decade of SMC proteins. Genes Dev. 19, 1269–1287 (2005)

    Article  Google Scholar 

  4. Hauf, S. & Watanabe, Y. Kinetochore orientation in mitosis and meiosis. Cell 119, 317–327 (2004)

    Article  CAS  Google Scholar 

  5. Uhlmann, F. Chromosome cohesion and separation: from men and molecules. Curr. Biol. 13, R104–R114 (2003)

    Article  CAS  Google Scholar 

  6. Lee, J. Y. & Orr-Weaver, T. L. The molecular basis of sister-chromatid cohesion. Annu. Rev. Cell Dev. Biol. 17, 753–777 (2001)

    Article  CAS  Google Scholar 

  7. Watanabe, Y. Shugoshin: guardian spirit at the centromere. Curr. Opin. Cell Biol. 17, 590–595 (2005)

    Article  CAS  Google Scholar 

  8. Kitajima, T. S., Kawashima, S. A. & Watanabe, Y. The conserved kinetochore protein shugoshin protects centromeric cohesion during meiosis. Nature 427, 510–517 (2004)

    Article  ADS  CAS  Google Scholar 

  9. Rabitsch, K. P. et al. Two fission yeast homologs of Drosophila Mei-S332 are required for chromosome segregation during meiosis I and II. Curr. Biol. 14, 287–301 (2004)

    Article  CAS  Google Scholar 

  10. Marston, A. L., Tham, W. H., Shah, H. & Amon, A. A genome-wide screen identifies genes required for centromeric cohesion. Science 303, 1367–1370 (2004)

    Article  ADS  CAS  Google Scholar 

  11. Katis, V. L., Galova, M., Rabitsch, K. P., Gregan, J. & Nasmyth, K. Maintenance of cohesin at centromeres after meiosis I in budding yeast requires a kinetochore-associated protein related to MEI-S332. Curr. Biol. 14, 560–572 (2004)

    Article  CAS  Google Scholar 

  12. Hamant, O. et al. A REC8-dependent plant shugoshin is required for maintenance of centromeric cohesion during meiosis and has no mitotic functions. Curr. Biol. 15, 948–954 (2005)

    Article  CAS  Google Scholar 

  13. Salic, A., Waters, J. C. & Mitchison, T. J. Vertebrate shugoshin links sister centromere cohesion and kinetochore microtubule stability in mitosis. Cell 118, 567–578 (2004)

    Article  CAS  Google Scholar 

  14. Kitajima, T. S., Hauf, S., Ohsugi, M., Yamamoto, T. & Watanabe, Y. Human Bub1 defines the persistent cohesion site along the mitotic chromosome by affecting Shugoshin localization. Curr. Biol. 15, 353–359 (2005)

    Article  CAS  Google Scholar 

  15. McGuinness, B. E., Hirota, T., Kudo, N. R., Peters, J.-M. & Nasmyth, K. Shugoshin prevents dissociation of cohesin from centromeres during mitosis in vertebrate cells. PLoS Biol. 3, e86 (2005)

    Article  Google Scholar 

  16. Sumara, I. et al. The dissociation of cohesin from chromosomes in prophase is regulated by Polo-like kinase. Mol. Cell 9, 515–525 (2002)

    Article  CAS  Google Scholar 

  17. Hauf, S. et al. Dissociation of cohesin from chromosome arms and loss of arm cohesion during early mitosis depends on phosphorylation of SA2. PLoS Biol. 3, e69 (2005)

    Article  Google Scholar 

  18. Losada, A., Hirano, M. & Hirano, T. Cohesin release is required for sister chromatid resolution, but not for condensin-mediated compaction, at the onset of mitosis. Genes Dev. 16, 3004–3016 (2002)

    Article  CAS  Google Scholar 

  19. Alexandru, G., Uhlmann, F., Mechtler, K., Poupart, M. & Nasmyth, K. Phosphorylation of the cohesin subunit Scc1 by Polo/Cdc5 kinase regulates sister chromatid separation in yeast. Cell 105, 459–472 (2001)

    Article  CAS  Google Scholar 

  20. Hornig, N. C. & Uhlmann, F. Preferential cleavage of chromatin-bound cohesin after targeted phosphorylation by Polo-like kinase. EMBO J. 23, 3144–3153 (2004)

    Article  CAS  Google Scholar 

  21. Lee, B. H. & Amon, A. Role of Polo-like kinase CDC5 in programming meiosis I chromosome segregation. Science 300, 482–486 (2003)

    Article  ADS  CAS  Google Scholar 

  22. Clyne, R. K. et al. Polo-like kinase Cdc5 promotes chiasmata formation and cosegregation of sister centromeres at meiosis I. Nature Cell Biol. 5, 480–485 (2003)

    Article  CAS  Google Scholar 

  23. Natsume, T. et al. A direct nanoflow liquid chromatography–tandem mass spectrometry system for interaction proteomics. Anal. Chem. 74, 4725–4733 (2002)

    Article  CAS  Google Scholar 

  24. Janssens, V. & Goris, J. Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem. J. 353, 417–439 (2001)

    Article  CAS  Google Scholar 

  25. Tang, Z., Sun, Y., Harley, S. E., Zou, H. & Yu, H. Human Bub1 protects centromeric sister-chromatid cohesion through Shugoshin during mitosis. Proc. Natl Acad. Sci. USA 101, 18012–18017 (2004)

    Article  ADS  CAS  Google Scholar 

  26. Vandre, D. D. & Wills, V. L. Inhibition of mitosis by okadaic acid: possible involvement of a protein phosphatase 2A in the transition from metaphase to anaphase. J. Cell Sci. 101, 79–91 (1992)

    CAS  PubMed  Google Scholar 

  27. Van Dolah, F. M. & Ramsdell, J. S. Okadaic acid inhibits a protein phosphatase activity involved in formation of the mitotic spindle of GH4 rat pituitary cells. J. Cell. Physiol. 152, 190–198 (1992)

    Article  CAS  Google Scholar 

  28. Jiang, W. & Hallberg, R. L. Isolation and characterization of par1+ and par2+: two Schizosaccharomyces pombe genes encoding B′ subunits of protein phosphatase 2A. Genetics 154, 1025–1038 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Le Goff, X. et al. The protein phosphatase 2A B′-regulatory subunit par1p is implicated in regulation of the S. pombe septation initiation network. FEBS Lett. 508, 136–142 (2001)

    Article  CAS  Google Scholar 

  30. Kinoshita, N., Ohkura, H. & Yanagida, M. Distinct, essential roles of type 1 and 2A protein phosphatases in control of the fission yeast cell division cycle. Cell 63, 405–415 (1990)

    Article  CAS  Google Scholar 

  31. Mailhes, J. B., Hilliard, C., Fuseler, J. W. & London, S. N. Okadaic acid, an inhibitor of protein phosphatase 1 and 2A, induces premature separation of sister chromatids during meiosis I and aneuploidy in mouse oocytes in vitro. Chromosome Res. 11, 619–631 (2003)

    Article  CAS  Google Scholar 

  32. Kiburz, B. M. et al. The core centromere and Sgo1 establish a 50-kb cohesin-protected domain around centromeres during meiosis I. Genes Dev. 19, 3017–3030 (2005)

    Article  CAS  Google Scholar 

  33. Clarke, A. S., Tang, T. T., Ooi, D. L. & Orr-Weaver, T. L. POLO kinase regulates the Drosophila centromere cohesion protein MEI-S332. Dev. Cell 8, 53–64 (2005)

    Article  CAS  Google Scholar 

  34. Janssens, V., Goris, J. & Van Hoof, C. PP2A: the expected tumor suppressor. Curr. Opin. Genet. Dev. 15, 34–41 (2005)

    Article  CAS  Google Scholar 

  35. Nakajima, H., Toyoshima-Morimoto, F., Taniguchi, E. & Nishida, E. Identification of a consensus motif for Plk (Polo-like kinase) phosphorylation reveals Myt1 as a Plk1 substrate. J. Biol. Chem. 278, 25277–25280 (2003)

    Article  CAS  Google Scholar 

  36. Hauf, S. et al. The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint. J. Cell Biol. 161, 281–294 (2003)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank S. Hauf for critically reading the manuscript. We also thank B. Akiyoshi for yeast two-hybrid screening of S. pombe Sgo1; K. Nasmyth for communicating unpublished results; J. M. Peters, E. Nishida, F. Ishikawa, Y. Nakatani, M. Sato and T. Toda for reagents; J. M. Peters, M. Ohsugi and M. Shimura for methods; and all members in our laboratory for their valuable discussion and help. K.I. was supported by a JSPS fellowship. This work was supported in part by the New Energy and Industrial Technology Development Organization (to T.N.), and by the Toray Science Foundation, Uehara Memorial Foundation, and a Grant-in-Aid for Specially Promoted Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to Y.W.). Author Contributions Experiments in Figs 1 –3 were mainly performed by T.S.K., Fig. 4 by K.I., and Figs 5 and 6 by T.S. and S.A.K. Mass spectrometry of hSgo1 immunoprecipitates was performed by S.I. and T.N. Experimental design, interpretation of data, and the preparation of the manuscript were mainly conducted by T.S.K., T.S., K.I. and Y.W.

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Authors and Affiliations

  1. Laboratory of Chromosome Dynamics, Institute of Molecular and Cellular Biosciences,

    Tomoya S. Kitajima, Takeshi Sakuno, Kei-ichiro Ishiguro, Shigehiro A. Kawashima & Yoshinori Watanabe

  2. Graduate Program in Biophysics and Biochemistry, University of Tokyo,

    Shigehiro A. Kawashima & Yoshinori Watanabe

  3. SORST, Japan Science and Technology Agency, Yayoi, Tokyo, 113-0032, Japan

    Takeshi Sakuno & Yoshinori Watanabe

  4. National Institute of Advanced Industrial Science and Technology, Biological Information Research Center, Aomi, Tokyo, 135-0064, Japan

    Shun-ichiro Iemura & Tohru Natsume

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Correspondence to Yoshinori Watanabe.

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This file contains Supplementary Table 1, Supplementary Figures 1–9, Supplementary Methods and additional references. (PDF 4449 kb)

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Kitajima, T., Sakuno, T., Ishiguro, Ki. et al. Shugoshin collaborates with protein phosphatase 2A to protect cohesin. Nature 441, 46–52 (2006). https://doi.org/10.1038/nature04663

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