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Nonhomologous end-joining of site-specific but not of radiation-induced DNA double-strand breaks is reduced in the presence of wild-type p53

Oncogene volume 24, pages 1663–1672 (2005)Cite this article

Abstract

Nonhomologous end-joining (NHEJ) of DNA double-strand breaks (DSBs) entails two principal mechanisms: modification of DNA ends prior to ligation (error-prone rejoining) or precise ligation without modification if the DNA ends are complementary (error-free repair). Error-prone rejoining is mutagenic, because it can lead to destruction of coding sequence or to chromosomal aberrations, and therefore must be tightly regulated. Previous studies on the role of the p53 tumor suppressor in the regulation of NHEJ have yielded conflicting results, but a rigorous analysis of NHEJ proficiency and fidelity in a purely chromosomal context has not been carried out. To this end, we created novel repair plasmid substrates that integrate into the genome. DSBs generated by the I-SceI endonuclease within these substrates were repaired by either error-prone rejoining or precise ligation. We found that the expression of wild-type p53 inhibited any repair-associated DNA sequence deletion, including a more than 250-fold inhibition of error-prone rejoining events compared to p53-null cells, while any promoting effect of p53 on precise ligation could not be directly evaluated. The role of p53 in NHEJ appeared to involve a direct transactivation-independent mechanism, possibly restricting DNA end-modification by blocking the annealing of single strands along flanking stretches of microhomology. The inhibition of error-prone rejoining by p53 did not apply to the rejoining of DSBs induced by ionizing radiation. In conclusion, our data suggest that p53 restricts the mutagenic effects of NHEJ without compromising repair proficiency or cell survival, thereby maintaining genomic stability.

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Acknowledgements

We thank Dr Fen Xia and the members of their labs for critical review of the manuscript, Drs Maria Jasin and Heiner Küpper for their generous contribution of plasmids, and Lena Schliecker for technical assistance. This work was supported by a grant from the Deutsche Krebshilfe (German Cancer Aid) to JDD (10-1843-Da) and a grant from the NCI/NIH to SNP (R01 CA58985).

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Authors and Affiliations

  1. Laboratory of Experimental Radiation Oncology, Department of Radiotherapy and Radiation Oncology, University Hospital Eppendorf, University of Hamburg, 20246, Hamburg, Germany

    Jochen Dahm-Daphi, Petra Hubbe, Fruzsina Horvath & Raafat A El-Awady

  2. Laboratory of Molecular and Cellular Radiation Biology, Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, 02129, USA

    Katie E Bouffard, Simon N Powell & Henning Willers

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  1. Jochen Dahm-Daphi
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Correspondence to Jochen Dahm-Daphi.

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Dahm-Daphi, J., Hubbe, P., Horvath, F. et al. Nonhomologous end-joining of site-specific but not of radiation-induced DNA double-strand breaks is reduced in the presence of wild-type p53. Oncogene 24, 1663–1672 (2005). https://doi.org/10.1038/sj.onc.1208396

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