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Molecular and Cellular Biology, March 2007, p. 1602-1613, Vol. 27, No. 5
0270-7306/07/$08.00+0     doi:10.1128/MCB.01956-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

RSC Mobilizes Nucleosomes To Improve Accessibility of Repair Machinery to the Damaged Chromatin{triangledown} ,{dagger}

Eun Yong Shim, Soo Jin Hong, Ji-Hyun Oum, Yvonne Yanez, Yu Zhang, and Sang Eun Lee*

Department of Molecular Medicine and Institute of Biotechnology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78245

Received 17 October 2006/ Returned for modification 6 November 2006/ Accepted 6 December 2006

Repair of DNA double-strand breaks (DSBs) protects cells and organisms, as well as their genome integrity. Since DSB repair occurs in the context of chromatin, chromatin must be modified to prevent it from inhibiting DSB repair. Evidence supports the role of histone modifications and ATP-dependent chromatin remodeling in repair and signaling of chromosome DSBs. The key questions are, then, what the nature of chromatin altered by DSBs is and how remodeling of chromatin facilitates DSB repair. Here we report a chromatin alteration caused by a single HO endonuclease-generated DSB at the Saccharomyces cerevisiae MAT locus. The break induces rapid nucleosome migration to form histone-free DNA of a few hundred base pairs immediately adjacent to the break. The DSB-induced nucleosome repositioning appears independent of end processing, since it still occurs when the 5'-to-3' degradation of the DNA end is markedly reduced. The tetracycline-controlled depletion of Sth1, the ATPase of RSC, or deletion of RSC2 severely reduces chromatin remodeling and loading of Mre11 and Yku proteins at the DSB. Depletion of Sth1 also reduces phosphorylation of H2A, processing, and joining of DSBs. We propose that RSC-mediated chromatin remodeling at the DSB prepares chromatin to allow repair machinery to access the break and is vital for efficient DSB repair.


* Corresponding author. Mailing address: Department of Molecular Medicine and Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 15355 Lambda Drive, San Antonio, TX 78245. Phone: (210) 567-7273. Fax: (210) 567-7269. E-mail: lees4{at}uthscsa.edu.

{triangledown} Published ahead of print on 18 December 2006.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.


Molecular and Cellular Biology, March 2007, p. 1602-1613, Vol. 27, No. 5
0270-7306/07/$08.00+0     doi:10.1128/MCB.01956-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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