Abstract
Repair of DNA double-strand breaks (DSBs) by homologous recombination (HR) requires that the 5'-terminated DNA strands are resected to generate single-stranded DNA overhangs. This process is initiated by a short-range resection catalyzed by the MRX (Mre11-Rad50-Xrs2) complex, which is followed by a long-range step involving the nucleases Exo1 and Dna2. Here we show that the Saccharomyces cerevisiae ATP-dependent chromatin-remodeling protein Chd1 participates in both short- and long-range resection by promoting MRX and Exo1 association with the DSB ends. Furthermore, Chd1 reduces histone occupancy near the DSB ends and promotes DSB repair by HR. All these functions require Chd1 ATPase activity, supporting a role for Chd1 in the opening of chromatin at the DSB site to facilitate MRX and Exo1 processing activities.
Highlights
DNA double-strand breaks (DSBs) are the most common cause of genomic instability, because their inaccurate repair can lead to chromosomal rearrangements
Eukaryotic genomes are compacted into chromatin, which restricts the access to DNA of the enzymes devoted to repair DNA DSBs
We examine the role in DSB repair of the ATP-dependent chromatin remodeler chromodomain-helicase-DNA-binding protein 1 (Chd1), which is frequently mutated in prostate cancer
Summary
Strain genotypes are listed in S1 Table. Strains JKM139, YMV45 and tGI354, used to detect DSB resection, DSB repair by SSA and DSB repair by ectopic recombination, respectively, were kindly provided by J. Cells were grown in YEP medium (1% yeast extract, 2% bactopeptone) supplemented with 2% glucose (YEPD), 2% raffinose (YEPR) or 2% raffinose and 3% galactose (YEPRG). Gene disruptions were generated by one-step PCR homology cassette amplification and standard yeast transformation method.
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