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Forced activation of Cdk1 via wee1 inhibition impairs homologous recombination

Oncogene volume 32, pages 3001–3008 (2013)Cite this article

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Abstract

In response to DNA breaks, the ‘DNA damage response’ provokes a cell cycle arrest to facilitate DNA repair. Recent findings have indicated that cells can respond to DNA damage throughout the cell cycle, except during mitosis. Specifically, various mitotic kinases, including Cdk1, Aurora A and Plk1, were shown to inactivate key DNA damage checkpoint proteins when cells enter mitosis. Aberrant activation of mitotic kinases during interphase could therefore modulate cellular responses to DNA damage. In this study, our aim was to determine how aberrant activation of Cdk1 affects the cellular responses to DNA damage. We used Wee1 inhibition, using MK-1775, to force Cdk1 activation, which did not cause cytotoxicity in non-transformed cells. Instead, it accelerated mitotic entry and caused radio sensitization in p53-defective cancer cells, but not in p53-proficient cancer cells. Interestingly, we showed that Wee1 inhibition leads to elevation of Cdk1 activity in interphase cells. When we subsequently analyzed DNA damage responses in cells with forced Cdk1 activation, we observed a marked reduction of 53BP1 at sites of DNA damage along with an increase in γ-H2AX staining after irradiation, indicative of defective DNA repair. Indeed, when DNA repair was analyzed using in vivo endonuclease-induced homologous recombination (HR) assays, compromised DNA repair after Wee1 inhibition was confirmed. This defect in HR was accompanied by increased phosphorylation of BRCA2 at the Cdk1 phosphorylation site S3291. Taken together, our results indicate that Wee1 inhibition leads to forced Cdk1 activation in interphase cells, which interferes with normal DNA damage responses.

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Acknowledgements

This work was supported by a NWO-VENI grant (916.76.062) to MATMVV. We thank Drs Maria Jasin, Jeff Parvin, Reuven Agami, James Ferrell Jr and Vera Gorbunova for generously supplying materials, Dr Gerben Vader for critically reading the manuscript and members of the Medical Oncology laboratory for helpful discussions.

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Authors and Affiliations

  1. Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands

    M Krajewska, A M Heijink, Y J W M Bisselink, R I Seinstra, E G E de Vries & M A T M van Vugt

  2. Department of Cardiology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands

    H H W Silljé

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  1. M Krajewska
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  2. A M Heijink
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Correspondence to M A T M van Vugt.

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Krajewska, M., Heijink, A., Bisselink, Y. et al. Forced activation of Cdk1 via wee1 inhibition impairs homologous recombination. Oncogene 32, 3001–3008 (2013). https://doi.org/10.1038/onc.2012.296

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