Centrosome amplification induced by DNA damage occurs during a prolonged G2 phase and involves ATM

Abstract

Centrosomes are the principal microtubule organising centres in somatic cells. Abnormal centrosome number is common in tumours and occurs after γ‐irradiation and in cells with mutations in DNA repair genes. To investigate how DNA damage causes centrosome amplification, we examined cells that conditionally lack the Rad51 recombinase and thereby incur high levels of spontaneous DNA damage. Rad51‐deficient cells arrested in G2 phase and formed supernumerary functional centrosomes, as assessed by light and serial section electron microscopy. This centrosome amplification occurred without an additional DNA replication round and was not the result of cytokinesis failure. G2‐to‐M checkpoint over‐ride by caffeine or wortmannin treatment strongly reduced DNA damage‐induced centrosome amplification. Radiation‐induced centrosome amplification was potentiated by Rad54 disruption. Gene targeting of ATM reduced, but did not abrogate, centrosome amplification induced by DNA damage in both the Rad51 and Rad54 knockout models, demonstrating ATM‐dependent and ‐independent components of DNA damage‐inducible G2‐phase centrosome amplification. Our data suggest DNA damage‐induced centrosome amplification as a mechanism for ensuring death of cells that evade the DNA damage or spindle assembly checkpoints.