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Nucleic Acids Research Advance Access originally published online on September 4, 2008
Nucleic Acids Research 2008 36(17):5678-5694; doi:10.1093/nar/gkn550
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Nucleic Acids Research, 2008, Vol. 36, No. 17 5678-5694
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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{gamma}-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin

Andrea Kinner, Wenqi Wu, Christian Staudt and George Iliakis*

Institute of Medical Radiation Biology, University of Duisburg-Essen Medical School, Hufelandstrasse 55, 45122 Essen, Germany

*To whom correspondence should be addressed. Tel: +49 201 7234152; Fax: +49 201 7235966; Email: georg.lliakis{at}uk-essen.de

Received June 27, 2008. Revised August 11, 2008. Accepted August 12, 2008.

DNA double-strand breaks (DSBs) are extremely dangerous lesions with severe consequences for cell survival and the maintenance of genomic stability. In higher eukaryotic cells, DSBs in chromatin promptly initiate the phosphorylation of the histone H2A variant, H2AX, at Serine 139 to generate {gamma}-H2AX. This phosphorylation event requires the activation of the phosphatidylinositol-3-OH-kinase-like family of protein kinases, DNA-PKcs, ATM, and ATR, and serves as a landing pad for the accumulation and retention of the central components of the signaling cascade initiated by DNA damage. Regions in chromatin with {gamma}-H2AX are conveniently detected by immunofluorescence microscopy and serve as beacons of DSBs. This has allowed the development of an assay that has proved particularly useful in the molecular analysis of the processing of DSBs. Here, we first review the role of {gamma}-H2AX in DNA damage response in the context of chromatin and discuss subsequently the use of this modification as a surrogate marker for mechanistic studies of DSB induction and processing. We conclude with a critical analysis of the strengths and weaknesses of the approach and present some interesting applications of the resulting methodology.


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