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Abstract

Segregation of homologous maternal and paternal centromeres to opposite poles during meiosis I depends on post-replicative crossing over between homologous non-sister chromatids, which creates chiasmata and therefore bivalent chromosomes. Destruction of sister chromatid cohesion along chromosome arms due to proteolytic cleavage of cohesin's Rec8 subunit by separase resolves chiasmata and thereby triggers the first meiotic division. This produces univalent chromosomes, the chromatids of which are held together by centromeric cohesin that has been protected from separase by shugoshin (Sgo1/MEI-S332) proteins. Here we show in both fission and budding yeast that Sgo1 recruits to centromeres a specific form of protein phosphatase 2A (PP2A). Its inactivation causes loss of centromeric cohesin at anaphase I and random segregation of sister centromeres at the second meiotic division. Artificial recruitment of PP2A to chromosome arms prevents Rec8 phosphorylation and hinders resolution of chiasmata. Our data are consistent with the notion that efficient cleavage of Rec8 requires phosphorylation of cohesin and that this is blocked by PP2A at meiosis I centromeres.

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Acknowledgements

The authors would like to thank K. Rabitsch, N. Kudo, C. Kraft, I. Steinmacher, R. Imre, K. Tanaka, R. Kittler, F. Stewart and A. Lorenz for help with experiments, as well as M. Yanagida, I. Poser, I. Hagan, K. Gull, K. Gould, C. Warren, F. Spencer, D. Virshup, W. Zachariae and Y. Watanabe for cell lines, strains and reagents. In addition we would like to thank members of the Nasmyth laboratory for discussions. J.G. was a recipient of the EMBO long-term fellowship. K.M. received funding from GenAU. E.O. was supported by a grant from the Austrian Science Foundation. Author Contributions Experiments were designed and data analysed and interpreted by C.R., V.K. and K.N. S. pombe strain construction, cytological and biochemical analysis, TAP purification, conventional ChIP experiments and in vitro binding assays were performed by C.R. S. cerevisiae strain construction and cytological analysis was performed by V.K. ChIP using S. pombe and S. cerevisiae oligonucleotide microarrays was performed by Y.K., S.M., T.I. and K.S. S. cerevisiae TAP purification was performed by M.P. HeLa TAP purification was performed by J.G. from a cell line with help from F.B. and L.P. Cytological analysis of HeLa and NIH3T3 cells was performed by B.C., using cell lines generated by I.M. and E.O. Generation and characterization of human αB56 antibody was performed by I.M. and E.O. Mass spectrometry was performed by K.M. The manuscript was written by V.K., C.R. and K.N. W.H. and M.G. provided technical assistance.

Author information

Author notes

    • Vittorio L. Katis
    • , Wolfgang Helmhart
    • , Bulent Cetin
    •  & Kim Nasmyth

    †Present address: Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK

    • Christian G. Riedel
    •  & Vittorio L. Katis

    *These authors contributed equally to this work

Affiliations

  1. Research Institute of Molecular Pathology (IMP), Dr. Bohr-Gasse 7, A-1030 Vienna, Austria

    • Christian G. Riedel
    • , Vittorio L. Katis
    • , Wolfgang Helmhart
    • , Marta Gálová
    • , Mark Petronczki
    • , Juraj Gregan
    • , Bulent Cetin
    • , Karl Mechtler
    •  & Kim Nasmyth
  2. Laboratory of Genome Structure and Function, Division of Gene Research, Center for Biological Resources and Informatics, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan

    • Yuki Katou
    • , Saori Mori
    •  & Katsuhiko Shirahige
  3. Research Center for Advanced Science and Technology, Mitsubishi Research Institute Inc., 2-3-6 Ohtemachi, Chiyoda-ku, Tokyo 100-8141, Japan

    • Takehiko Itoh
  4. Department of Medical Biochemistry, Max F. Perutz Laboratories, Vienna Biocenter, Medical University of Vienna, Dr. Bohr-Gasse 9/2, A-1030 Vienna, Austria

    • Ingrid Mudrak
    •  & Egon Ogris
  5. Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Pfotenhauerstrasse, 01307 Dresden, Germany

    • Laurence Pelletier
    •  & Frank Buchholz

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Corresponding author

Correspondence to Kim Nasmyth.

Supplementary information

PDF files

  1. 1.

    Supplementary Notes

    This file contains Supplementary Methods, Supplementary Tables 1 and 2, and Supplementary Figures 1–3, 6–10 and Supplementary Figure Legends.

  2. 2.

    Supplementary Figure 4

    Distribution of Sp_Rec8, Sp_Par1B’, Sp_Ppa2C, and Sp_Sgo1 on chromatin, using a high-density oligonucleotide array of Schizosaccharomyces pombe chromosome 2 and most of chromosome 3.

  3. 3.

    Supplementary Figure 5

    Distribution of Sp_Par1B’ on chromatin in Sp_sgo1Δ cells, using a high-density oligonucleotide array of S. pombe chromosome 2 and most of chromosome 3.

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DOI

https://doi.org/10.1038/nature04664

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