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The mitotic checkpoint complex binds a second CDC20 to inhibit active APC/C

Nature volume 517, pages 631634 (29 January 2015) | Download Citation

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Abstract

The spindle assembly checkpoint (SAC) maintains genomic stability by delaying chromosome segregation until the last chromosome has attached to the mitotic spindle. The SAC prevents the anaphase promoting complex/cyclosome (APC/C) ubiquitin ligase from recognizing cyclin B and securin by catalysing the incorporation of the APC/C co-activator, CDC20, into a complex called the mitotic checkpoint complex (MCC). The SAC works through unattached kinetochores generating a diffusible ‘wait anaphase’ signal1,2 that inhibits the APC/C in the cytoplasm, but the nature of this signal remains a key unsolved problem. Moreover, the SAC and the APC/C are highly responsive to each other: the APC/C quickly targets cyclin B and securin once all the chromosomes attach in metaphase, but is rapidly inhibited should kinetochore attachment be perturbed3,4. How this is achieved is also unknown. Here, we show that the MCC can inhibit a second CDC20 that has already bound and activated the APC/C. We show how the MCC inhibits active APC/C and that this is essential for the SAC. Moreover, this mechanism can prevent anaphase in the absence of kinetochore signalling. Thus, we propose that the diffusible ‘wait anaphase’ signal could be the MCC itself, and explain how reactivating the SAC can rapidly inhibit active APC/C.

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Acknowledgements

We are grateful to T. Matsusaka for developing infrared-dye-conjugated ubiquitylation substrates, to A. Musacchio, W. Earnshaw, T. Kiyomitsu and M. Yanagida for reagents, and to A. Musacchio and members of our laboratory for critical discussions. This work was supported by a project and a programme grant from Cancer Research UK to J.P. J.P. acknowledges core funding to the Gurdon Institute from the Wellcome Trust and CR UK.

Author information

Affiliations

  1. The Gurdon Institute and Department of Zoology, Tennis Court Road, Cambridge CB2 1QN, UK

    • Daisuke Izawa
    •  & Jonathon Pines

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Contributions

Experiments were designed by D.I. and J.P., carried out by D.I., and analysed by D.I. and J.P.; D.I. and J.P. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jonathon Pines.

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https://doi.org/10.1038/nature13911

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