Centrosome duplication is critical for cell division, and genome instability can result if duplication is not restricted to a single round per cell cycle. Centrosome duplication is controlled in part by CP110, a centriolar protein that positively regulates centriole duplication while restricting centriole elongation and ciliogenesis. Maintenance of normal CP110 levels is essential, as excessive CP110 drives centrosome over-duplication and suppresses ciliogenesis, whereas its depletion inhibits centriole amplification and leads to highly elongated centrioles and aberrant assembly of cilia in growing cells1,2. CP110 levels are tightly controlled, partly through ubiquitination by the ubiquitin ligase complex SCFcyclin F during G2 and M phases of the cell cycle3. Here, using human cells, we report a new mechanism for the regulation of centrosome duplication that requires USP33, a deubiquitinating enzyme that is able to regulate CP110 levels. USP33 interacts with CP110 and localizes to centrioles primarily in S and G2/M phases, the periods during which centrioles duplicate and elongate. USP33 potently and specifically deubiquitinates CP110, but not other cyclin-F substrates. USP33 activity antagonizes SCFcyclin F-mediated ubiquitination and promotes the generation of supernumerary centriolar foci, whereas ablation of USP33 destabilizes CP110 and thereby inhibits centrosome amplification and mitotic defects. To our knowledge, we have identified the first centriolar deubiquitinating enzyme whose expression regulates centrosome homeostasis by countering cyclin-F-mediated destruction of a key substrate. Our results point towards potential therapeutic strategies for inhibiting tumorigenesis associated with centrosome amplification.
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We thank C. Hajdu, G. Miller, L. Chiriboga, H. Court, C. Zambirinis, E. Bekes, L. Taylor, S. Krauter, M. Philips and D. Bar-Sagi for help with staining and analysis of pancreatic cancer samples. We thank F. Liang, Y. Deng, C. Petzold and K. Dancel for help with correlative electron microscopy. We thank J. Jastrab and S. Vijayakumar for initial experiments pertaining to the USP20–CP110 interaction. We thank J. Pollack for providing pancreatic cell lines. We thank G. Wu, S. Shenoy, R. Baer and T. Huang for providing plasmids. We thank members of the Dynlacht laboratory and T. Huang for advice. B.D.D. was supported by National Institutes of Health grant 5R01HD069647-02 and March of Dimes grant FY11-432. M.P. is an Investigator with the Howard Hughes Medical Institute.
The authors declare no competing financial interests.
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Li, J., D’Angiolella, V., Seeley, E. et al. USP33 regulates centrosome biogenesis via deubiquitination of the centriolar protein CP110. Nature 495, 255–259 (2013) doi:10.1038/nature11941
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