Letter | Published:

Rec8 phosphorylation and recombination promote the step-wise loss of cohesins in meiosis

Naturevolume 441pages532536 (2006) | Download Citation

Subjects

Abstract

During meiosis, cohesins—protein complexes that hold sister chromatids together—are lost from chromosomes in a step-wise manner1. Loss of cohesins from chromosome arms is necessary for homologous chromosomes to segregate during meiosis I. Retention of cohesins around centromeres until meiosis II is required for the accurate segregation of sister chromatids. Here we show that phosphorylation of the cohesin subunit Rec8 contributes to step-wise cohesin removal. Our data further implicate two other key regulators of meiotic chromosome segregation, the cohesin protector Sgo1 and meiotic recombination in bringing about the step-wise loss of cohesins and thus the establishment of the meiotic chromosome segregation pattern. Understanding the interplay between these processes should provide insight into the events underlying meiotic chromosome mis-segregation, the leading cause of miscarriages and mental retardation in humans.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Marston, A. L. & Amon, A. Meiosis: cell-cycle controls shuffle and deal. Nature Rev. Mol. Cell Biol. 5, 983–997 (2004)

  2. 2

    Buonomo, S. B. et al. Division of the nucleolus and its release of Cdc14 during anaphase of meiosis I depends on Separase, Spo12, and Slk19. Dev. Cell 4, 727–739 (2003)

  3. 3

    Tang, T. T., Bickel, S. E., Young, L. M. & Orr-Weaver, T. L. Maintenance of sister-chromatid cohesion at the centromere by the Drosophila MEI-S332 protein. Genes Dev. 12, 3843–3856 (1998)

  4. 4

    Kerrebrock, A. W., Moore, D. P., Wu, J. S. & Orr-Weaver, T. L. MEI-S332, a Drosophila protein required for sister-chromatid cohesion, can localize to meiotic centromere regions. Cell 83, 247–256 (1995)

  5. 5

    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)

  6. 6

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

  7. 7

    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)

  8. 8

    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)

  9. 9

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

  10. 10

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

  11. 11

    Nasmyth, K. & Haering, C. H. The structure and function of SMC and kleisin complexes. Annu. Rev. Biochem. 74, 595–648 (2005)

  12. 12

    Yu, H. G. & Koshland, D. Chromosome morphogenesis: Condensin-dependent cohesin removal during meiosis. Cell 123, 397–407 (2005)

  13. 13

    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)

  14. 14

    Lee, B. H. & Amon, A. Role of Polo-like kinase Cdc5 in programming meiosis I chromosome segregation. Science 300, 482–486 (2003)

  15. 15

    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)

  16. 16

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

  17. 17

    Klein, F. et al. A central role for cohesins in sister chromatid cohesion, formation of axial elements, and recombination during yeast meiosis. Cell 98, 91–103 (1999)

  18. 18

    Buonomo, S. B. et al. Disjunction of homologous chromosomes in meiosis I depends on proteolytic cleavage of the meiotic cohesin Rec8 by Separase. Cell 103, 387–398 (2000)

  19. 19

    Keeney, S., Giroux, C. N. & Kleckner, N. Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88, 375–384 (1997)

  20. 20

    Bergerat, A. et al. An atypical topoisomerase II from Archaea with implications for meiotic recombination. Nature 386, 414–417 (1997)

  21. 21

    Toth, A. et al. Functional genomics identifies monopolin: A kinetochore protein required for segregation of homologs during meiosis I. Cell 103, 1155–1168 (2000)

  22. 22

    Alani, E., Padmore, R. & Kleckner, N. Analysis of wild-type and rad50 mutants of yeast suggests an intimate relationship between meiotic chromosome synapsis and recombination. Cell 61, 419–436 (1990)

  23. 23

    Bishop, D. K., Park, D., Xu, L. & Kleckner, N. Dmc1: A meiosis-specific yeast homolog of E. coli RecA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell 69, 439–456 (1992)

  24. 24

    Shonn, M. A., Murray, A. L. & Murray, A. W. Spindle checkpoint component Mad2 contributes to biorientation of homologous chromosomes. Curr. Biol. 13, 1979–1984 (2003)

  25. 25

    Shonn, M. A., McCarroll, R. & Murray, A. W. Requirement of the spindle checkpoint for proper chromosome segregation in budding yeast meiosis. Science 289, 300–303 (2000)

  26. 26

    Cohen-Fix, O., Peters, J. M., Kirschner, M. W. & Koshland, D. Anaphase initiation in Saccharomyces cerevisiae is controlled by the APC-dependent degradation of the anaphase inhibitor Pds1p. Genes Dev. 10, 3081–3093 (1996)

  27. 27

    Nairz, K. & Klein, F. mre11S—a yeast mutation that blocks double-strand-break processing and permits nonhomologous synapsis in meiosis. Genes Dev. 11, 2272–2290 (1997)

  28. 28

    Visintin, R. et al. The phosphatase Cdc14 triggers mitotic exit by reversal of CDK-dependent phosphorylation. Mol. Cell 2, 709–718 (1998)

Download references

Acknowledgements

We thank F. Lewitter for help with the Polo kinase substrate consensus sequence analysis, L.-S. Ee for technical assistance, K. Nasmyth and Y. Watanabe for communication of results before publication and B. Lee, A. Marston and members of the Amon Lab for input and critical reading of the manuscript. A.A. is an Investigator of the Howard Hughes Medical Institute. This research was supported by an NIH grant to A.A. and a NSF pre-doctoral fellowship to G.A.B. Author Contributions G.A.B., B.M.K. and A.A. participated in the design of experiments. G.A.B. and B.M.K. performed all non-MS experiments. Y.Z., J.-E.K. and F.W. designed and performed all MS analysis and analysed MS results.

Author information

Affiliations

  1. Center for Cancer Research, Howard Hughes Medical Institute, Massachusetts Institute of Technology, E17-233 40 Ames Street, Massachusetts, 02139, Cambridge, USA

    • Gloria A. Brar
    • , Brendan M. Kiburz
    •  & Angelika Amon
  2. Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Building 56-787a, Massachusetts, 02139-4307, Cambridge, USA

    • Yi Zhang
    • , Ji-Eun Kim
    •  & Forest White

Authors

  1. Search for Gloria A. Brar in:

  2. Search for Brendan M. Kiburz in:

  3. Search for Yi Zhang in:

  4. Search for Ji-Eun Kim in:

  5. Search for Forest White in:

  6. Search for Angelika Amon in:

Competing interests

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

Corresponding author

Correspondence to Angelika Amon.

Supplementary information

  1. Supplementary Figure 1

    Example of a MS/MS spectrum. (PDF 29 kb)

  2. Supplementary Figure 2

    Coverage of the Rec8 in the various cell cycle arrest. (PDF 92 kb)

  3. Supplementary Figure 3

    Mutation of the phosphorylation sites in Rec8 to alanine interferes with progression through meiosis I. (PDF 66 kb)

  4. Supplementary Figure 4

    Mutations of phosphorylation or cleavage sites of Rec8 interferes with chromosome segregation. (PDF 38 kb)

  5. Supplementary Figure 5

    Metaphase I cells lacking Pds1 accumulate in rec8-17A mutants. (PDF 128 kb)

  6. Supplementary Figure 6

    Recombination allows Rec8 cleavage in rec8-17A and rec8-29A mutants. (PDF 135 kb)

  7. Supplementary Figure 7

    Rec8-17A cleavage in cells expressing a catalytically dead version of SPO11. (PDF 92 kb)

  8. Supplementary Figure 8

    Deletion of SPO11 abolishes step-wise loss of meiotic cohesion. (PDF 73 kb)

  9. Supplementary Figure 9

    Depletion of Sgo1 in spo11-Y135F, rad50S or dmc1Δ mek1Δ mutants allows proper meiosis II chromosome segregation in Sgo1-depleted cells. (PDF 27 kb)

  10. Supplementary Figure 10

    Depletion of Sgo1 partially alleviates the need for Rec8 phosphorylation and Cdc5 in Rec8 cleavage and anaphase I entry. (PDF 142 kb)

  11. Supplementary Figure 11

    Depletion of Sgo1 allows Rec8 removal from chromosomes in Cdc5-depleted cells. (PDF 832 kb)

  12. Supplementary Figure 12

    Rec8 phosphorylation on S136 and S521 is regulated during meiosis. (PDF 40 kb)

  13. Supplementary Table 1

    Cdc5 independent phosphorylation sites. (DOC 80 kb)

  14. Supplementary Table 2

    CDC5-dependent phosphorylation sites (PDF 43 kb)

  15. Supplementary Notes

    This file contains the Supplementary Methods, Supplementary Figure Legends, Supplementary Table 3 and additional references. (DOC 152 kb)

About this article

Publication history

Received

Accepted

Published

Issue Date

DOI

https://doi.org/10.1038/nature04794

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.