Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Shugoshin–PP2A counteracts casein-kinase-1-dependent cleavage of Rec8 by separase

Abstract

During meiosis, the cohesin complexes that maintain sister chromatid cohesion are lost in a stepwise manner1,2. At meiosis I the cohesin subunit Rec8 is cleaved only along the chromosome arms; until meiosis II it is protected at centromeres by the action of shugoshin (Sgo1)–protein phosphatase 2A (PP2A)3,4,5. Although this regulation hypothetically involves phosphorylation that is antagonized by Sgo1–PP2A, the kinase and substrate that are responsible are as yet unknown6,7. Using a genetic screen for 'anti-shugoshin', we identify Hhp2, an orthologue of casein kinase 1δ/ɛ (CK1), as a factor required for Rec8 cleavage in fission yeast. We show that CK1, rather than a Polo-like kinase that is widely believed to do so, acts as the cohesin kinase to promote this cleavage during meiosis. Crucially, forced localization of excess Hhp2 at the pericentromeric region abrogates the ability of Sgo1–PP2A to protect centromeric Rec8. Thus, our studies prove the key notion that the balance between Rec8 phosphorylation and its dephosphorylation by Sgo1–PP2A regulates the step-wise loss of chromosomal cohesion in meiosis.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Overproduction of Rec8 in mitosis causes chromosome segregation defects in hhp2Δ cells.
Figure 2: CK1, but not Plo1, is important for Rec8 phosphorylation and cleavage in meiosis.
Figure 3: Identification of CK1-dependent phosphorylation sites required for Rec8 cleavage.
Figure 4: Phosphorylation of Rec8 by CK1 is essential for cleavage by separase.
Figure 5: Forced localization of CK1 removes centromeric Rec8 and induces precocious centromeric separation before meiosis II.

Similar content being viewed by others

References

  1. Petronczki, M., Siomos, M. F. & Nasmyth, K. Un ménage à quatre: the molecular biology of chromosome segregation in meiosis. Cell 112, 423–440 (2003).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  4. Kitajima, T. S. et al. Shugoshin collaborates with protein phosphatase 2A to protect cohesin. Nature 441, 46–52 (2006).

    Article  CAS  Google Scholar 

  5. Riedel, C. G. et al. Protein phosphatase 2A protects centromeric sister chromatid cohesion during meiosis I. Nature 441, 53–61 (2006).

    Article  CAS  Google Scholar 

  6. Brar, G. A. et al. Rec8 phosphorylation and recombination promote the step-wise loss of cohesins in meiosis. Nature 441, 532–536 (2006).

    Article  CAS  Google Scholar 

  7. Kudo, N. R. et al. Role of cleavage by separase of the Rec8 kleisin subunit of cohesin during mammalian meiosis I. J. Cell Sci. 122, 2686–2698 (2009).

    Article  CAS  Google Scholar 

  8. Peters, J. M., Tedeschi, A. & Schmitz, J. The cohesin complex and its roles in chromosome biology. Genes Dev. 22, 3089–3114 (2008).

    Article  CAS  Google Scholar 

  9. Hirano, T. At the heart of the chromosome: SMC proteins in action. Nature Rev. Mol. Cell Biol. 7, 311–322 (2006).

    Article  CAS  Google Scholar 

  10. 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 

  11. 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 

  12. Alexandru, G., Uhlmann, F., Mechtler, K., Poupart, M. A. & 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 

  13. 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 

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

    Article  CAS  Google Scholar 

  15. Dhillon, N. & Hoekstra, M. F. Characterization of two protein kinases from Schizosaccharomyces pombe involved in the regulation of DNA repair. EMBO J. 13, 2777–2788 (1994).

    Article  CAS  Google Scholar 

  16. Kearney, P. H., Ebert, M. & Kuret, J. Molecular cloning and sequence analysis of two novel fission yeast casein kinase-1 isoforms. Biochem. Biophys. Res. Commun. 203, 231–236 (1994).

    Article  CAS  Google Scholar 

  17. Petronczki, M. et al. Monopolar attachment of sister kinetochores at meiosis I requires casein kinase 1. Cell 126, 1049–1064 (2006).

    Article  CAS  Google Scholar 

  18. Mata, J., Lyne, R., Burns, G. & Bahler, J. The transcriptional program of meiosis and sporulation in fission yeast. Nature Genet. 32, 143–147 (2002).

    Article  CAS  Google Scholar 

  19. Ohkura, H., Hagan, I. M. & Glover, D. M. The conserved Schizosaccharomyces pombe kinase plo1, required to form a bipolar spindle, the actin ring, and septum, can drive septum formation in G1 and G2 cells. Genes Dev. 9, 1059–1073 (1995).

    Article  CAS  Google Scholar 

  20. Flotow, H. et al. Phosphate groups as substrate determinants for casein kinase I action. J. Biol. Chem. 265, 14264–14269 (1990).

    CAS  PubMed  Google Scholar 

  21. Bustos, V. H. et al. Generation of protein kinase Ck1α mutants which discriminate between canonical and non-canonical substrates. Biochem. J. 391, 417–424 (2005).

    Article  CAS  Google Scholar 

  22. Zhou, F. F., Xue, Y., Chen, G. L. & Yao, X. GPS: a novel group-based phosphorylation predicting and scoring method. Biochem. Biophys. Res. Commun. 325, 1443–1448 (2004).

    Article  CAS  Google Scholar 

  23. Nakayama, J., Rice, J. C., Strahl, B. D., Allis, C. D. & Grewal, S. I. Role of histone H3 lysine 9 methylation in epigenetic control of heterochromatin assembly. Science 292, 110–113 (2001).

    Article  CAS  Google Scholar 

  24. Yamagishi, Y., Sakuno, T., Shimura, M. & Watanabe, Y. Heterochromatin links to centromeric protection by recruiting shugoshin. Nature 455, 251–255 (2008).

    Article  CAS  Google Scholar 

  25. Sakuno, T., Tada, K. & Watanabe, Y. Kinetochore geometry defined by cohesion within the centromere. Nature 458, 852–858 (2009).

    Article  CAS  Google Scholar 

  26. Kitajima, T. S., Miyazaki, Y., Yamamoto, M. & Watanabe, Y. Rec8 cleavage by separase is required for meiotic nuclear divisions in fission yeast. EMBO J. 22, 5643–5653 (2003).

    Article  CAS  Google Scholar 

  27. Katis, V. L. et al. Rec8 phosphorylation by casein kinase 1 and Cdc7-Dbf4 kinase regulates cohesin cleavage by separase during meiosis. Dev. Cell 18, 397–409 (2010).

    Article  CAS  Google Scholar 

  28. Bähler, J. et al. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast 14, 943–951 (1998).

    Article  Google Scholar 

  29. Sato, M., Dhut, S. & Toda, T. New drug-resistant cassettes for gene disruption and epitope tagging in Schizosaccharomyces pombe. Yeast 22, 583–591 (2005).

    Article  CAS  Google Scholar 

  30. Nabeshima, K. et al. Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis1 is implicated in force balance in metaphase bipolar spindle. Mol. Biol. Cell 9, 3211–3225 (1998).

    Article  CAS  Google Scholar 

  31. Yamamoto, A. & Hiraoka, Y. Monopolar spindle attachment of sister chromatids is ensured by two distinct mechanisms at the first meiotic division in fission yeast. EMBO J. 22, 2284–2296 (2003).

    Article  CAS  Google Scholar 

  32. Watanabe, Y., Yokobayashi, S., Yamamoto, M. & Nurse, P. Pre-meiotic S phase is linked to reductional chromosome segregation and recombination. Nature 409, 359–363 (2001).

    Article  CAS  Google Scholar 

  33. Harigaya, Y. et al. Selective elimination of messenger RNA prevents an incidence of untimely meiosis. Nature 442, 45–50 (2006).

    Article  CAS  Google Scholar 

  34. Yokobayashi, S. & Watanabe, Y. The kinetochore protein Moa1 enables cohesion-mediated monopolar attachment at meiosis I. Cell 123, 803–817 (2005).

    Article  CAS  Google Scholar 

  35. Takayama, Y. et al. Biphasic incorporation of centromeric histone CENP-A in fission yeast. Mol. Biol. Cell 19, 682–690 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Silke Hauf for critically reading the manuscript; Iain Hagan and the Yeast Genetic Resource Center (YGRC) for yeast strains; and all the members of our laboratory for their valuable support and discussion. We also thank Kim Nasmyth and Wolfgang Zachariae for communicating unpublished results. This work was supported in part by Special Coordination Funds for Promoting Science and Technology (to T.S.) and the Global COE Program (Integrative Life Science Based on the Study of Biosignaling Mechanisms) and a Grant-in-Aid for Scientific research on Priority Areas (to K.T. and T.S.) and for Specially Promoted Research (to Y.W.), MEXT, Japan.

Author information

Authors and Affiliations

Authors

Contributions

Experimental design and interpretation of data were conducted by all authors. T.I., K.T. and T.S. performed experiments. Y.W. planned the project and wrote the paper with input from the co-authors.

Corresponding author

Correspondence to Yoshinori Watanabe.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 2622 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ishiguro, T., Tanaka, K., Sakuno, T. et al. Shugoshin–PP2A counteracts casein-kinase-1-dependent cleavage of Rec8 by separase. Nat Cell Biol 12, 500–506 (2010). https://doi.org/10.1038/ncb2052

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb2052

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing