Histone acetylation by Trrap–Tip60 modulates loading of repair proteins and repair of DNA double-strand breaks

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Abstract

DNA is packaged into chromatin, a highly compacted DNA–protein complex; therefore, all cellular processes that use the DNA as a template, including DNA repair, require a high degree of coordination between the DNA-repair machinery and chromatin modification/remodelling, which regulates the accessibility of DNA in chromatin. Recent studies have implicated histone acetyltransferase (HAT) complexes and chromatin acetylation in DNA repair; however, the precise underlying mechanism remains poorly understood1,2. Here, we show that the HAT cofactor Trrap and Tip60 HAT bind to the chromatin surrounding sites of DNA double-strand breaks (DSBs) in vivo. Trrap depletion impairs both DNA-damage-induced histone H4 hyperacetylation and accumulation of repair molecules at sites of DSBs, resulting in defective homologous recombination (HR) repair, albeit with the presence of a functional ATM-dependent DNA-damage signalling cascade. Importantly, the impaired loading of repair proteins and the defect in DNA repair in Trrap-deficient cells can be counteracted by chromatin relaxation, indicating that the DNA-repair defect that was observed in the absence of Trrap is due to impeded chromatin accessibility at sites of DNA breaks. Thus, these data reveal that cells may use the same basic mechanism involving HAT complexes to regulate distinct cellular processes, such as transcription and DNA repair.

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Figure 1: Impaired homologous recombination repair of DSBs in cells lacking Trrap.
Figure 2: Histone H4 acetylation and occupancy of Tip60 near DSBs.
Figure 3: Trrap depletion impairs recruitment of DNA-repair proteins to sites of DNA breaks.
Figure 4: Normal DNA-damage sensing or signalling in cells lacking Trrap.
Figure 5: Defect in HR repair and impaired recruitment of repair proteins in cells lacking Trrap may be improved by chromatin relaxation.

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Acknowledgements

We thank M. Jasin (Memorial Sloan-Kettering Cancer Center, New York, NY) for hprt–DRGFP and pCBASce vectors, D. Livingston (Harvard Medical School, Boston, MA) for anti-BRCA1 antibodies, J. Chen (Mayo Clinic, Rochester, MN) for anti-53BP1 and anti-Mdc1 antibodies, L. Tora (Institut de Genetique et de Biologie Moleculaire et Cellulaire, IGBMC, Illkirch, France) for anti-TRRAP antibodies and communicating unpublished results, and S. Kochbin (Institut Albert Bonniot, La Tronche, France) for Tip60 constructs. We also thank V. Shukla for help in cell culture and M.-P. Cros for the maintenance of mouse colonies and assistance in collecting blastocysts. Further thanks are due to J. Hall, B. Sylla and M. Finkbeiner for critical reading of the manuscript and helpful discussions. We are grateful to J. Cheney and M. Renaud for editing the manuscript. R.M. is supported by a PhD fellowship from la Ligue Nationale (Française) Contre le Cancer. J.I.L. is supported by a postdoctoral fellowship from the International Agency for Research on Cancer (IARC) and by an EMBO long-term fellowship. This work was supported by the Association pour la Recherche sur le Cancer (ARC), France and the Association for International Cancer Research (AICR), UK.

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Correspondence to Zdenko Herceg.

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