title: RSC mobilizes nucleosomes to improve accessibility of repair machinery to the damaged chromatin.
abstract: 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.
authors.affiliation: Department of Molecular Medicine and Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 15355 Lambda Drive, San Antonio, TX 78245, USA.
authors.forename: Eun Yong
authors.initials: EY
authors.lastname: Shim
authors.forename: Soo Jin
authors.initials: SJ
authors.lastname: Hong
authors.forename: Ji-Hyun
authors.initials: JH
authors.lastname: Oum
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Yanez
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Sang Eun
authors.initials: SE
authors.lastname: Lee
mesh_terms: D002843:Chromatin / Q000235:genetics* / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D015825:Chromosomes, Fungal; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004271:DNA, Fungal; D004268:DNA-Binding Proteins / Q000378:metabolism*; D009707:Nucleosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D002843:Chromatin; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D009707:Nucleosomes; C103337:RSC complex, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors