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p53 modulates homologous recombination at I-SceI-induced double-strand breaks through cell-cycle regulation

Oncogene volume 32, pages 968–975 (2013)Cite this article

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Abstract

Inhibition of homologous recombination (HR) is believed to be a transactivation-independent function of p53 that protects from genetic instability. Misrepair by HR can lead to genetic alterations such as translocations, duplications, insertions and loss of heterozygosity, which all bear the risk of driving oncogenic transformation. Regulation of HR by wild-type p53 (wtp53) should prevent these genomic rearrangements. Mutation of p53 is a frequent event during carcinogenesis. In particular, dominant-negative mutants inhibiting wtp53 expressed from the unperturbed allel can drive oncogenic transformation by disrupting the p53-dependent anticancer barrier. Here, we asked whether the hot spot mutants R175H and R273H relax HR control in p53-proficient cells. Utilizing an I-SceI-based reporter assay, we observed a moderate (1.5 × ) stimulation of HR upon expression of the mutant proteins in p53-proficient CV-1, but not in p53-deficient H1299 cells. Importantly, the stimulatory effect was exactly paralleled by an increase in the number of HR competent S- and G2-phase cells, which can well explain the enhanced recombination frequencies. Furthermore, the impact on HR exerted by the transactivation domain double-mutant L22Q/W23S and mutant R273P, both of which were reported to regulate HR independently of G1-arrest execution, is also exactly mirrored by cell-cycle behavior. These results are in contrast to previous concepts stating that the transactivation-independent impact of p53 on HR is a general phenomenon valid for replication-associated and also for directly induced double-strand break. Our data strongly suggest that the latter is largely mediated by cell-cycle regulation, a classical transactivation-dependent function of p53.

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Acknowledgements

We thank Dr Tom Misteli for the pDsRed-I-SceI-GR vector. This work was supported by a grant from the German Cancer Aid (Deutsche Krebshilfe No. 107889 to JDD and No. 107980 to ID).

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Authors and Affiliations

  1. Laboratory of Radiobiology and Experimental Radiooncology, University Medical Center Hamburg–Eppendorf, Hamburg, Germany

    T Rieckmann, M Kriegs, L Nitsch, K Hoffer, S Kocher, E Dikomey & J Dahm-Daphi

  2. Heinrich Pette-Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany

    G Rohaly & I Dornreiter

  3. Institute of Radiobiology and Molecular Radiation Oncology, Particle Therapy Center, Philipps-University, Marburg, Germany

    S Kocher & J Dahm-Daphi

  4. Department of Radiotherapy and Radiooncology, University Medical Center Hamburg–Eppendorf, Hamburg, Germany

    C Petersen

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  1. T Rieckmann
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Correspondence to J Dahm-Daphi.

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Rieckmann, T., Kriegs, M., Nitsch, L. et al. p53 modulates homologous recombination at I-SceI-induced double-strand breaks through cell-cycle regulation. Oncogene 32, 968–975 (2013). https://doi.org/10.1038/onc.2012.123

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