Anaphase initiation is regulated by antagonistic ubiquitination and deubiquitination activities (original) (raw)

• Article
• Published: 19 April 2007
• Michael Rape3 na1 nAff7,
• Viji M. Draviam3,4,
• Grzegorz Nalepa2,
• Mathew E. Sowa2,
• Xiaolu L. Ang2,
• E. Robert McDonald III1,
• Mamie Z. Li1,
• Gregory J. Hannon5,
• Peter K. Sorger3,4,
• Marc W. Kirschner3,
• J. Wade Harper2 &
• …
• Stephen J. Elledge1

Nature volume 446, pages 876–881 (2007)Cite this article

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Abstract

The spindle checkpoint prevents chromosome mis-segregation by delaying sister chromatid separation until all chromosomes have achieved bipolar attachment to the mitotic spindle. Its operation is essential for accurate chromosome segregation, whereas its dysregulation can contribute to birth defects and tumorigenesis. The target of the spindle checkpoint is the anaphase-promoting complex (APC), a ubiquitin ligase that promotes sister chromatid separation and progression to anaphase. Using a short hairpin RNA screen targeting components of the ubiquitin-proteasome pathway in human cells, we identified the deubiquitinating enzyme USP44 (ubiquitin-specific protease 44) as a critical regulator of the spindle checkpoint. USP44 is not required for the initial recognition of unattached kinetochores and the subsequent recruitment of checkpoint components. Instead, it prevents the premature activation of the APC by stabilizing the APC-inhibitory Mad2–Cdc20 complex. USP44 deubiquitinates the APC coactivator Cdc20 both in vitro and in vivo, and thereby directly counteracts the APC-driven disassembly of Mad2–Cdc20 complexes (discussed in an accompanying paper). Our findings suggest that a dynamic balance of ubiquitination by the APC and deubiquitination by USP44 contributes to the generation of the switch-like transition controlling anaphase entry, analogous to the way that phosphorylation and dephosphorylation of Cdk1 by Wee1 and Cdc25 controls entry into mitosis.

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Acknowledgements

We thank S. Taylor, H. Yu, W. Earnshaw and J. Jin for gifts of reagents; M. Vidal for providing access to their BioRobot platform; S. Lyman and R. King for communicating unpublished results and assistance with the development of the Taxol screening assay; C. Shamu for access to the ICCB-Longwood screening facilities; S. Reddy for helpful comments throughout the course of the work; and T. Westbrook and A. Smogorzewska for their critical reading of the manuscript. F.S. is a fellow of the Helen Hay Whitney Foundation. M.R. is a Human Frontiers Science Program Long-Term Fellow. The siRNA and ICCB-Longwood resources used were funded in part by a NCI grant (T. Mitchison). M.E.S. is an American Cancer Society Postdoctoral Fellow. X.L.A. is an NIH pre-doctoral fellow. M.W.K. thanks the National Institute of General Medical Sciences for its support for the grant Cell Cycle Regulation. This work was supported by grants from NIH and DOD to S.J.E. and by grants from the NIH to J.W.H. S.J.E. is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

1. Michael Rape
   Present address: Present address: Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, California 94720-3202, USA.,
2. Frank Stegmeier and Michael Rape: These authors contributed equally to this work.

Authors and Affiliations

1. Department of Genetics, Howard Hughes Medical Institute, Harvard Partners Center for Genetics and Genomics, and,
   Frank Stegmeier, E. Robert McDonald III, Mamie Z. Li & Stephen J. Elledge
2. Department of Pathology, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA,
   Grzegorz Nalepa, Mathew E. Sowa, Xiaolu L. Ang & J. Wade Harper
3. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA,
   Michael Rape, Viji M. Draviam, Peter K. Sorger & Marc W. Kirschner
4. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA,
   Viji M. Draviam & Peter K. Sorger
5. Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, New York 11724, USA,
   Gregory J. Hannon

Authors

1. Frank Stegmeier
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2. Michael Rape
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3. Viji M. Draviam
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4. Grzegorz Nalepa
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5. Mathew E. Sowa
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6. Xiaolu L. Ang
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7. E. Robert McDonald III
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8. Mamie Z. Li
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9. Gregory J. Hannon
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10. Peter K. Sorger
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11. Marc W. Kirschner
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12. J. Wade Harper
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13. Stephen J. Elledge
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Corresponding authors

Correspondence toMarc W. Kirschner, J. Wade Harper or Stephen J. Elledge.

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Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Figures 1-11 with Legends, and Supplementary Tables 2 and 3. (PDF 4063 kb)

Supplementary Table 1

This file contains Supplementary Table 1. This file contains detailed information on the Ubiquitin-Proteasome Pathway (UPP) shRNA library. (XLS 526 kb)

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Stegmeier, F., Rape, M., Draviam, V. et al. Anaphase initiation is regulated by antagonistic ubiquitination and deubiquitination activities.Nature 446, 876–881 (2007). https://doi.org/10.1038/nature05694

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• Received: 13 December 2006
• Accepted: 19 February 2007
• Issue Date: 19 April 2007
• DOI: https://doi.org/10.1038/nature05694

Editorial Summary

Cell division control

During cell division the spindle checkpoint ensures that chromosome segregation is delayed until all chromosomes are properly attached to the mitotic spindle. Two papers now identify a new regulatory mechanism that controls the spindle checkpoint. This involves the fine-tuned ubiquitination and de-ubiquitination of a coactivator of the anaphase promoting complex APC/C to regulate the timing of APC/C activation and thereby the onset of anaphase.

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