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:

APCCdc20 promotes exit from mitosis by destroying the anaphase inhibitor Pds1 and cyclin Clb5

Nature volume 402, pages 203–207 (1999)Cite this article

Abstract

Ubiquitin-mediated proteolysis due to the anaphase-promoting complex/cyclosome (APC/C) is essential for separation of sister chromatids, requiring degradation of the anaphase inhibitor Pds1, and for exit from mitosis, requiring inactivation of cyclin B Cdk1 kinases1. Exit from mitosis in yeast involves accumulation of the cyclin kinase inhibitor Sic1 as well as cyclin proteolysis mediated by APC/C bound by the activating subunit Cdh1/Hct1 (APCCdh1)2,3. Both processes require the Cdc14 phosphatase, whose release from the nucleolus during anaphase causes dephosphorylation and thereby activation of Cdh1 and accumulation of another protein, Sic1 (refs 4,5,6,7). We do not know what determines the release of Cdc14 and enables it to promote Cdk1 inactivation, but it is known to be dependent on APC/C bound by Cdc20 (APCCdc20) (ref. 4). Here we show that APCCdc20 allows activation of Cdc14 and promotes exit from mitosis by mediating proteolysis of Pds1 and the S phase cyclin Clb5 in the yeast Saccharomyces cerevisiae. Degradation of Pds1 is necessary for release of Cdc14 from the nucleolus, whereas degradation of Clb5 is crucial if Cdc14 is to overwhelm Cdk1 and activate its foes (Cdh1 and Sic1). Remarkably, cells lacking both Pds1 and Clb5 can proliferate in the complete absence of Cdc20.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Deletion of CLB5 bypasses lethality of cdc20 pds1 null mutants.
Figure 2: Characterization of cdc20 and cdc20 clb5 mutants.
Figure 3: Cdc14 delocalizes into nucleus in cdc20 pds1 mutants.
Figure 4: APCCdc20 coordinates ‘sister separation’ and ‘exit from mitosis’ during the cell cycle.

Similar content being viewed by others

References

  1. Morgan,D. O. Regulation of the APC and the exit from mitosis. Nature Cell Biol. 1, E47–E53 (1999).

    Article  CAS  Google Scholar 

  2. Visintin,R., Prinz,S. & Amon,A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science 278, 460–463 (1997).

    Article  ADS  CAS  Google Scholar 

  3. Schwab,M., Lutum,A. S. & Seufert,W. Yeast Hct1 is a regulator of Clb2 cyclin proteolysis. Cell 90, 683–693 (1997).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  5. Jaspersen,S. L., Charles,J. F. & Morgan,D. O. Inhibitory phosphorylation of the APC regulator Hct1 is controlled by the kinase Cdc28 and the phosphatase Cdc14. Curr. Biol. 9, 227–236 (1999).

    Article  CAS  Google Scholar 

  6. Shou,W. et al. Exit from mitosis is triggered by Tem1-dependent release of the protein phosphatase Cdc14 from nucleolar RENT complex. Cell 97, 233–244 (1999).

    Article  CAS  Google Scholar 

  7. Visintin,R., Hwang,E. S. & Amon,A. Cfi1 prevents premature exit from mitosis by anchoring Cdc14 phosphatase in the nucleolus. Nature 398, 818–823 (1999).

    Article  ADS  CAS  Google Scholar 

  8. Yamamoto,A., Guacci,V. & Koshland,D. Pds1p, an inhibitor of anaphase in budding yeast, plays a critical role in the APC and checkpoint pathway(s). J. Cell Biol. 133, 99–110 (1996).

    Article  CAS  Google Scholar 

  9. Shirayama,M., Zachariae,W., Ciosk,R. & Nasmyth,K. The Polo-like kinase Cdc5p and the WD-repeat protein Cdc20p/Fizzy are regulators and substrates of the anaphase promoting complex in Saccharomyces cerevisiae. EMBO J. 17, 1336–1349 (1998).

    Article  CAS  Google Scholar 

  10. Lim,H. H., Goh,P. Y. & Surana,U. Cdc20 is essential for the cyclosome-mediated proteolysis of both Pds1 and Clb2 during M phase in budding yeast. Curr. Biol. 8, 231–234 (1998).

    Article  CAS  Google Scholar 

  11. Novák,B. et al. Finishing the cell cycle. J. Theor. Biol. 199, 223–233 (1999).

    Article  Google Scholar 

  12. Mumberg,D., Mueller,R. & Funk,M. Regulatable promoters of Saccharomyces cerevisiae: comparison of transcriptional activity and their use for heterologous expression. Nucleic Acids Res. 22, 5767–5768 (1994).

    Article  CAS  Google Scholar 

  13. Burns, N. et al. Large-scale analysis of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae. Genes Dev. 8, 1087–1105 (1994).

    Article  CAS  Google Scholar 

  14. Riley,J. et al. A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones. Nucleic Acids Res. 18, 2887–2890 (1990).

    Article  CAS  Google Scholar 

  15. Schwob,E., Böhm,T., Mendenhall,M. D. & Nasmyth,K. The B-type cyclin kinase inhibitor p40SIC1 controls the G1 to S transition in S. cerevisiae. Cell 79, 233–244 (1994).

    Article  CAS  Google Scholar 

  16. 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).

    Article  CAS  Google Scholar 

  17. de Beus,E., Brockenbrough,J. S., Hong,B. & Aris,J. P. Yeast NOP2 encodes an essential nucleolar protein with homology to a human proliferation marker. J. Cell Biol. 127, 1799–1813 (1994).

    Article  CAS  Google Scholar 

  18. Tinker-Kulberg,R. L. & Morgan,D. O. Pds1 and Esp1 control both anaphase and mitotic exit in normal cells and after DNA damage. Genes Dev. 13, 1936–1949 (1999).

    Article  CAS  Google Scholar 

  19. Cohen-Fix,O. & Koshland,D. Pds1p of budding yeast has dual roles: inhibition of anaphase initiation and regulation of mitotic exit. Genes Dev. 13, 1950–1959 (1999).

    Article  CAS  Google Scholar 

  20. Surana,U. et al. Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. EMBO J. 12, 1969–1978 (1993).

    Article  CAS  Google Scholar 

  21. Ciosk,R. et al. An Esp1/Pds1 complex regulates loss of sister chromatid cohesion at the metaphase to anaphase transition in yeast. Cell 93, 1067–1076 (1998).

    Article  CAS  Google Scholar 

  22. Skowyra,D. et al. F-box proteins are receptors that recruit phosphorylated substrates to the SCF ubiquitin-ligase complex. Cell 91, 209–219 (1997).

    Article  CAS  Google Scholar 

  23. Feldman,R. M., Correll,C. C., Kaplan,K. B. & Deshaies,R. J. A complex of Cdc4p, Skp1p, and Cdc53p/Cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p. Cell 91, 221–230 (1997).

    Article  CAS  Google Scholar 

  24. Verma,R. et al. Phosphorylation of Sic1p by G1 CDK required for its degradation and entry into S phase. Science 278, 455–460 (1997).

    Article  ADS  CAS  Google Scholar 

  25. Zachariae,W., Schwab,M., Nasmyth,K. & Seufert,W. Control of cyclin ubiquitination by CDK-regulated binding of Hct1 to the anaphase promoting complex. Science 282, 1721–1724 (1998).

    Article  ADS  CAS  Google Scholar 

  26. Epstein,C. B. & Cross,F. R. CLB5: a novel B cyclin from budding yeast with a role in S phase. Genes Dev. 6, 1695–1706 (1992).

    Article  CAS  Google Scholar 

  27. Schwob,E. & Nasmyth,K. CLB5 and CLB6, a new pair of B cyclins involved in DNA replication in Saccharomyces cerevisiae. Genes Dev. 7, 1160–1175 (1993).

    Article  CAS  Google Scholar 

  28. Sherr,C. J. Mammalian G1 cyclins. Cell 73, 1059–1065 (1993).

    Article  CAS  Google Scholar 

  29. Sigrist,S., Jacobs,H., Stratmann,R. & Lehner,C. F. Exit from mitosis is regulated by Drosophila fizzy and the sequential destruction of cyclins A, B and B3. EMBO J. 14, 4827–4838 (1995).

    Article  CAS  Google Scholar 

  30. Wach,A. et al. Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae. Yeast 13, 1065–1075 (1997).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank J. Charles, D. Morgan, A. Amon and E. Schwob for sharing unpublished results; E. Kramer and J.-M. Peters for technical help; A. Amon, J. Aris, L. Huber, L. Guarente, D. Kellogg, and J. Kilmartin for gifts of antibodies; G. Ammerer for the transposon library; and F. Cross, Y. Kikuchi and W. Zachariae for comments on this manuscript.

Author information

Authors and Affiliations

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

    Masaki Shirayama, Attila Tóth, Marta Gálová & Kim Nasmyth

Authors
  1. Masaki Shirayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Attila Tóth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marta Gálová
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kim Nasmyth
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kim Nasmyth.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shirayama, M., Tóth, A., Gálová, M. et al. APCCdc20 promotes exit from mitosis by destroying the anaphase inhibitor Pds1 and cyclin Clb5. Nature 402, 203–207 (1999). https://doi.org/10.1038/46080

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/46080

This article is cited by

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.

Search

Advanced 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