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Roles of ATM and NBS1 in chromatin structure modulation and DNA double-strand break repair

Nature Cell Biology volume 9, pages 683–690 (2007)Cite this article

Abstract

We developed a novel system to create DNA double-strand breaks (DSBs) at defined endogenous sites in the human genome, and used this system to detect protein recruitment and loss at and around these breaks by chromatin immunoprecipitation (ChIP). The detection of human ATM protein at site-specific DSBs required functional NBS1 protein, ATM kinase activity and ATM autophosphorylation on Ser 1981. DSB formation led to the localized disruption of nucleosomes, a process that depended on both functional NBS1 and ATM. These two proteins were also required for efficient recruitment of the repair cofactor XRCC4 to DSBs, and for efficient DSB repair. These results demonstrate the functional importance of ATM kinase activity and phosphorylation in the response to DSBs, and support a model in which ordered chromatin structure changes that occur after DNA breakage depend on functional NBS1 and ATM, and facilitate DNA DSB repair.

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Figure 1: Characterization of I-PpoI-mediated DSB formation and damage response.
Figure 2: ATM dimers are not detected at DSB sites.
Figure 3: Detection, chromatin modulation and repair of a site-specific DSB.
Figure 4: Histone loss from DSB sites and DSB repair are dependent on NBS1 and ATM.
Figure 5: Distribution of damage response proteins around a site-specific DSB.

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Acknowledgements

We thank D. Woods and M. Reis for excellent technical assistance. This work was supported by grants from the National Institutes of Health (CA71387 and CA21765 to M.B.K. and CA48022 to R.J.M., Jr.) and by the American Lebanese Syrian Associated Charities (ALSAC) of the St. Jude Children's Research Hospital.

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Authors and Affiliations

  1. Department of Oncology, St. Jude Children's Research Hospital, Memphis, 38105, TN, USA

    Elijahu Berkovich & Michael B. Kastan

  2. Departments of Pathology and of Genome Sciences, University of Washington, Box 357705, Seattle, 98195–7705, WA, USA

    Raymond J. Monnat Jr.

Authors
  1. Elijahu Berkovich
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  2. Raymond J. Monnat Jr.
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  3. Michael B. Kastan
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Correspondence to Michael B. Kastan.

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Supplementary figures S1, S2, S3, S4 and S5 and Supplementary Table S1 (PDF 1729 kb)

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Berkovich, E., Monnat, R. & Kastan, M. Roles of ATM and NBS1 in chromatin structure modulation and DNA double-strand break repair. Nat Cell Biol 9, 683–690 (2007). https://doi.org/10.1038/ncb1599

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