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

Nature Cell Biology volume 12, pages 500–506 (2010)Cite this article

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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.

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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.

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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 We also thank Kim Nasmyth and Wolfgang Zachariae for communicating unpublished results.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.

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Authors and Affiliations

  1. Laboratory of Chromosome Dynamics, Institute of Molecular and Cellular Biosciences, University of Tokyo, Yayoi, 113-0032, Tokyo, Japan

    Tadashi Ishiguro, Koichi Tanaka, Takeshi Sakuno & Yoshinori Watanabe

  2. Graduate Program in Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Yayoi, 113-0032, Tokyo, Japan

    Tadashi Ishiguro & Yoshinori Watanabe

  3. Promotion of Independence for Young Investigators, University of Tokyo, Yayoi, 113-0032, Tokyo, Japan

    Takeshi Sakuno

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  1. Tadashi Ishiguro
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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.

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Correspondence to Yoshinori Watanabe.

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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

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