p53-Driven apoptosis limits centrosome amplification and genomic instability downstream of NPM1 phosphorylation

Abstract

Chromosome loss or gain is associated with a large number of solid cancers, providing genomic plasticity and thus adaptability to cancer cells1,2. Numerical centrosome abnormalities arising from centrosome over-duplication or failed cytokinesis are a recognized cause of aneuploidy3,4. In higher eukaryotic cells, the centrosome duplicates only once per cell cycle to ensure the formation of a bipolar mitotic spindle that orchestrates the balanced distribution of the sister chromatids to the respective daughter cells5. Here we delineate the events that allow abnormal centrosome duplication, resulting in mitotic errors and incorrect chromosome segregation in cells with sustained cyclin-dependent kinase (CDK) activity. We have identified NPM1 as a substrate for CDK6 activated by the Kaposi's sarcoma herpesvirus (KSHV) D-type cyclin and shown that p53-driven apoptosis occurs downstream of NPM1 phosphorylation as a checkpoint mechanism that prevents accumulation of cells with supernumerary centrosomes. Our findings provide evidence that abnormal chromosome segregation in KSHV-infected cells is a direct consequence of NPM1 phosphorylation and predict that genomic instability is an inevitable consequence of latent KSHV infection.

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Figure 1: K-cyclin-driven abnormal centrosome duplication is independent of CDK2 activation.
Figure 2: NPM1 is a substrate for K-cyclin–CDK6 and its phosphorylation is essential for cyclin-driven centrosome duplication.
Figure 3: Cyclin expression cooperates with p53 loss in centrosome amplification.
Figure 4: Pro-apoptotic signalling limits K-cyclin driven centrosome amplification.
Figure 5: K-cyclin drives abnormal chromosome segregation in p53-negative backgrounds.

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Acknowledgements

This work was funded by a grant from Cancer Research UK. We thank Jeffrey Vieira for permission to use rKSHV and Dimitris Lagos and David J. Blackbourn for the generous gift of rKSHV BCBL-1 cells. We are grateful to Jonathan Pines and Kenji Fukasawa for providing essential reagents.

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M.E.C. designed and performed the experiments; A.K. and P.C provided crucial support and materials for the KESTREL screen; N.M. was responsible for the mass spectrometry data; H.P. assisted with the confocal and time-lapse microscopy; S.M. co-ordinated the research; M.E.C. and S.M. wrote the manuscript.

Corresponding author

Correspondence to Maria Emanuela Cuomo.

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The authors declare no competing financial interests.

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Cuomo, M., Knebel, A., Morrice, N. et al. p53-Driven apoptosis limits centrosome amplification and genomic instability downstream of NPM1 phosphorylation. Nat Cell Biol 10, 723–730 (2008). https://doi.org/10.1038/ncb1735

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