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How APC/C–Cdc20 changes its substrate specificity in mitosis

Nature Cell Biology volume 13, pages 223–233 (2011)Cite this article

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A Corrigendum to this article was published on 03 May 2011

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Abstract

Progress through mitosis requires that the right protein be degraded at the right time. One ubiquitin ligase, the anaphase-promoting complex or cyclosome (APC/C) targets most of the crucial mitotic regulators by changing its substrate specificity throughout mitosis. The spindle assembly checkpoint (SAC) acts on the APC/C co-activator, Cdc20 (cell division cycle 20), to block the degradation of metaphase substrates (for example, cyclin B1 and securin), but not others (for example, cyclin A). How this is achieved is unclear. Here we show that Cdc20 binds to different sites on the APC/C depending on the SAC. Cdc20 requires APC3 and APC8 to bind and activate the APC/C when the SAC is satisfied, but requires only APC8 to bind the APC/C when the SAC is active. Moreover, APC10 is crucial for the destruction of cyclin B1 and securin, but not cyclin A. We conclude that the SAC causes Cdc20 to bind to different sites on the APC/C and this alters APC/C substrate specificity.

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Figure 1: APC11 is required to degrade all model substrates.
Figure 2: APC3 is required to degrade both SAC-sensitive and SAC-insensitive substrates.
Figure 3: APC3 is required to bind Cdc20 only when the SAC is satisfied.
Figure 4: APC10 is required to recruit SAC-sensitive substrates.
Figure 5: A point mutation in APC8 is sufficient to reduce the binding of Cdc20 in prometaphase.
Figure 6: A point mutation in APC8 prevents cyclin A degradation in prometaphase.
Figure 7: The Cdc20-binding site on APC8 is also required in metaphase.

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  • 31 March 2011

    In the version of this article initially published online and in print, there were some data errors in table 1. These errors have been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank J-M. Peters for the anti-phospho APC1 antibody and S. Taylor for the HeLa-FRT cell line. We are grateful to D. Barford for discussing results before publication and all the members of our laboratory for comments and criticisms. D.I. was supported by a fellowship from the Japanese Society for the Promotion of Science and by the Association for International Cancer Research (AICR). The work was supported by core funding to the Gurdon Institute from the Wellcome Trust and Cancer Research UK, by a project grant from the AICR and a programme grant from Cancer Research UK to J.P.

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

    Daisuke Izawa & Jonathon Pines

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  1. Daisuke Izawa
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D.I. and J.P. designed and interpreted the experiments; D.I. carried out and analysed all the experiments; D.I. and J.P. wrote the paper.

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Correspondence to Jonathon Pines.

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Izawa, D., Pines, J. How APC/C–Cdc20 changes its substrate specificity in mitosis. Nat Cell Biol 13, 223–233 (2011). https://doi.org/10.1038/ncb2165

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