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J. Biol. Chem., Vol. 282, Issue 38, 27693-27701, September 21, 2007

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Dual Chromatin Remodeling Roles for RSC during DNA Double Strand Break Induction and Repair at the Yeast MAT Locus^*

Nicholas A. Kent¹, Anna L. Chambers², and Jessica A. Downs²³

From the Genetics Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, United Kingdom and the Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, United Kingdom

DNA double strand breaks (DSBs) are potentially serious chromosomal lesions. However, cells sometimes deliberately cleave their own DNA to facilitate certain chromosomal processes, and there is much interest in how such self-inflicted breaks are effectively managed. Eukaryotic DSBs occur in the context of chromatin and the RSC chromatin-remodeling ATPase complex has been shown to promote DSB repair at the budding yeast MAT locus DSB, created by the HO endonuclease during mating type switching. We show that the role of RSC at MAT is highly specialized. The Rsc1p subunit of RSC directs nucleosome sliding immediately after DSB creation at both MAT and generally and is required for efficient DNA damage-induced histone H2A phosphorylation and strand resection during repair by homologous recombination. However, the Rsc2p and Rsc7p subunits are additionally required to set up a basal MAT locus structure. This RSC-dependent chromatin structure at MAT ensures accessibility to the HO endonuclease. The RSC complex therefore has chromatin remodeling roles both before and after DSB induction at MAT, promoting both DNA cleavage and subsequent repair.

Received for publication, June 7, 2007 , and in revised form, July 20, 2007.

^* This work was supported by departmental grants (to N. A. K.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–3.

² Present address: Medical Research Council Genome Damage and Stability Centre, University of Sussex, Brighton BN1 9RQ, United Kingdom.

³ A Jenner Fellow of the Lister Institute of Preventative Medicine.

¹ To whom correspondence should be addressed: Cardiff School of Biosciences, Cardiff University, Museum Ave., Cardiff CF10 3US, United Kingdom. E-mail: kentn{at}cardiff.ac.uk.

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