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KAP-1 phosphorylation regulates CHD3 nucleosome remodeling during the DNA double-strand break response

Nature Structural & Molecular Biology volume 18, pages 831–839 (2011)Cite this article

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Abstract

KAP-1 poses a substantial barrier to DNA double-strand break (DSB) repair within heterochromatin that is alleviated by ATM-dependent KAP-1 phosphorylation (pKAP-1). Here we address the mechanistic consequences of pKAP-1 that promote heterochromatic DSB repair and chromatin relaxation. KAP-1 function involves autoSUMOylation and recruitment of nucleosome deacetylation, methylation and remodeling activities. Although heterochromatin acetylation or methylation changes were not detected, radiation-induced pKAP-1 dispersed the nucleosome remodeler CHD3 from DSBs and triggered concomitant chromatin relaxation; pKAP-1 loss reversed these effects. Depletion or inactivation of CHD3, or ablation of its interaction with KAP-1SUMO1, bypassed pKAP-1's role in repair. Though KAP-1 SUMOylation was unaffected after irradiation, CHD3 dissociated from KAP-1SUMO1 in a pKAP-1–dependent manner. We demonstrate that KAP-1Ser824 phosphorylation generates a motif that directly perturbs interactions between CHD3′s SUMO-interacting motif and SUMO1, dispersing CHD3 from heterochromatin DSBs and enabling repair.

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Figure 1: Heterochromatic DSB repair and DSB-induced nucleosome spacing alterations require sustained ATM activity and KAP-1 phosphorylation.
Figure 2: Ionizing radiation reduces CHD3 retention at sites of damage after ionizing radiation in an ATM-dependent, rapid and reversible manner.
Figure 3: Depletion or catalytic mutation of CHD3 alleviates the need for ATM activity or KAP-1 phosphorylation in DSB repair.
Figure 4: Ablating KAP-1 SUMOylation alleviates the need for pKAP-1 in DSB repair, and CHD3 interactions with KAP-1SUMO, but not overall KAP-1 SUMOylation, are altered after ionizing radiation.
Figure 5: pKAP-1 peptides disrupt interactions between SUMO1 and the SIM domain of CHD3 in vitro.

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Acknowledgements

We thank O.S. Gabrielsen and T. Sæther (University of Oslo) for the CHD3 construct, A. Oliver (University of Sussex) for helpful structural biology discussions and Y. Ziv (University of Tel Aviv) for advice on MNase assays. A.A.G. was supported by a grant from the Association for International Cancer Research (AICR). The P.A.J. lab is funded by the Medical Research Council (UK), AICR, the Wellcome Research Trust and the Department of Health Radiation Protection Programme (UK).

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  1. Genome Damage and Stability Centre, University of Sussex, East Sussex, UK

    Aaron A Goodarzi, Thomas Kurka & Penelope A Jeggo

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  1. Aaron A Goodarzi
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  3. Penelope A Jeggo
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Contributions

A.A.G. and T. K. conducted the experiments. A.A.G. and P.A.J. coauthored the manuscript and conceived of and designed the study.

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Correspondence to Aaron A Goodarzi or Penelope A Jeggo.

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Goodarzi, A., Kurka, T. & Jeggo, P. KAP-1 phosphorylation regulates CHD3 nucleosome remodeling during the DNA double-strand break response. Nat Struct Mol Biol 18, 831–839 (2011). https://doi.org/10.1038/nsmb.2077

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