Abstract
Repair of chromosome double-strand breaks (DSBs) is central to cell survival and genome integrity. Nonhomologous end joining (NHEJ) is the major cellular repair pathway that eliminates chromosome DSBs. Here we report our genetic screen that identified Rsc8 and Rsc30, subunits of the Saccharomyces cerevisiae chromatin remodeling complex RSC, as novel NHEJ factors. Deletion of RSC30 gene or the C-terminal truncation of RSC8 impairs NHEJ of a chromosome DSB created by HO endonuclease in vivo. rsc30Delta maintains a robust level of homologous recombination and the damage-induced cell cycle checkpoints. By chromatin immunoprecipitation, we show recruitment of RSC to a chromosome DSB with kinetics congruent with its involvement in NHEJ. Recruitment of RSC to a DSB depends on Mre11, Rsc30, and yKu70 proteins. Rsc1p and Rsc2p, two other RSC subunits, physically interact with yKu80p and Mre11p. The interaction of Rsc1p with Mre11p appears to be vital for survival from genotoxic stress. These results suggest that chromatin remodeling by RSC is important for NHEJ.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Cell Cycle
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Chromatin / metabolism
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Chromatin Assembly and Disassembly*
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Chromosomal Proteins, Non-Histone / chemistry
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Chromosomal Proteins, Non-Histone / metabolism
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DNA Damage*
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DNA Repair*
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DNA, Fungal / metabolism*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Deoxyribonucleases, Type II Site-Specific / metabolism
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Endodeoxyribonucleases / genetics
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Endodeoxyribonucleases / metabolism
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Exodeoxyribonucleases / genetics
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Exodeoxyribonucleases / metabolism
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Genes, Fungal / genetics
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Genetic Complementation Test
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Molecular Sequence Data
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Phenotype
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Protein Binding
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Protein Subunits / chemistry
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Protein Subunits / genetics
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Protein Subunits / metabolism
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Recombination, Genetic
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Saccharomyces cerevisiae / cytology
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Transcription Factors / chemistry
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Transcription Factors / genetics
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Transcription Factors / metabolism*
Substances
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Chromatin
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Chromosomal Proteins, Non-Histone
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DNA, Fungal
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DNA-Binding Proteins
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Protein Subunits
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RSC complex, S cerevisiae
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RSC1 protein, S cerevisiae
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RSC2 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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YKU70 protein, S cerevisiae
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high affinity DNA-binding factor, S cerevisiae
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Endodeoxyribonucleases
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Exodeoxyribonucleases
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MRE11 protein, S cerevisiae
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HO protein, S cerevisiae
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SCEI protein, S cerevisiae
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Deoxyribonucleases, Type II Site-Specific