title: Chromatin remodelling complex RSC promotes base excision repair in chromatin of Saccharomyces cerevisiae.
abstract: The base excision repair (BER) pathway is a conserved DNA repair system required to maintain genomic integrity and prevent mutagenesis in all eukaryotic cells. Nevertheless, how BER operates in vivo (i.e. in the context of chromatin) is poorly understood. We have investigated the role of an essential ATP-dependent chromatin remodelling (ACR) complex RSC (Remodels the Structure of Chromatin) in BER of intact yeast cells. We show that depletion of STH1, the ATPase subunit of RSC, causes enhanced sensitivity to the DNA alkylating agent methyl methanesulfonate (MMS) and results in a substantial inhibition of BER, at the GAL1 locus and in the genome overall. Consistent with this observation, the DNA in chromatin is less accessible to micrococcal nuclease digestion in the absence of RSC. Quantitative PCR results indicate that repair deficiency in STH1 depleted cells is not due to changes in the expression of BER genes. Collectively, our data indicates the RSC complex promotes efficient BER in chromatin. These results provide, for the first time, a link between ATP-dependent chromatin remodelling and BER in living cells.
authors.affiliation: Biochemistry and Biophysics, School of Molecular Biosciences, Washington State University, Pullman, WA 99164-7520, USA.
authors.forename: Wioletta
authors.initials: W
authors.lastname: Czaja
authors.affiliation: Biochemistry and Biophysics, School of Molecular Biosciences, Washington State University, Pullman, WA 99164-7520, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Mao
authors.affiliation: Biochemistry and Biophysics, School of Molecular Biosciences, Washington State University, Pullman, WA 99164-7520, USA. Electronic address: smerdon@wsu.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Smerdon
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002843:Chromatin / Q000235:genetics* / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D019175:DNA Methylation; D004260:DNA Repair / Q000187:drug effects; D004271:DNA, Fungal / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005686:Galactokinase / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D016681:Genome, Fungal; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D000477:Alkylating Agents; D018797:Cell Cycle Proteins; D002843:Chromatin; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C103337:RSC complex, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D008741:Methyl Methanesulfonate; C524219:GAL1 protein, S cerevisiae; D005686:Galactokinase; C077392:STH1 protein, S cerevisiae
keywords: Alkylated DNA damage; MMS; Nucleosome; SWI/SNF