Abstract
The temporal control of mitotic protein degradation remains incompletely understood. In particular, it is unclear why the mitotic checkpoint prevents the anaphase-promoting complex/cyclosome (APC/C)-mediated degradation of cyclin B and securin in early mitosis, but not cyclin A1,2,3. Here, we show that another APC/C substrate, NIMA-related kinase 2A (Nek2A), is also destroyed in pro-metaphase in a checkpoint-independent manner and that this depends on an exposed carboxy-terminal methionine–arginine (MR) dipeptide tail. Truncation of the Nek2A C terminus delays its degradation until late mitosis, whereas Nek2A C-terminal peptides interfere with APC/C activity in an MR-dependent manner. Most importantly, we show that Nek2A binds directly to the APC/C, also in an MR-dependent manner, even in the absence of the adaptor protein Cdc20. As similar C-terminal dipeptide tails promote direct association of Cdc20, Cdh1 and Apc10–Doc1 with core APC/C subunits, we propose that this sequence also allows a substrate, Nek2A, to directly bind the APC/C. Thus, although Cdc20 is required for the degradation of Nek2A, it is not required for its recruitment and this renders its degradation insensitive to the mitotic checkpoint.
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Acknowledgements
The authors thank J. Pines and R. Patel for helpful comments on the manuscript; J.-M. Peters, P. Clarke and J. Gannon for providing reagents; and T. Hunt and H. Mahbubani for access to the Cancer Research UK Clare Hall Laboratories Xenopus colony. This work was supported by grants to A.M.F. from the Association of International Cancer Research, the Wellcome Trust, the BBSRC and Cancer Research UK. H.Y. and Y.K. were supported by the Marie Curie Cancer Care and the Association of International Cancer Research. C.L. is an MRC Career Development Fellow and is also supported by Cancer Research UK.
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Hayes, M., Kimata, Y., Wattam, S. et al. Early mitotic degradation of Nek2A depends on Cdc20-independent interaction with the APC/C. Nat Cell Biol 8, 607–614 (2006). https://doi.org/10.1038/ncb1410
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DOI: https://doi.org/10.1038/ncb1410
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