DNA Damage-Induced Nucleosome Depletion Enhances Homology Search Independently of Local Break Movement

To determine whether double-strand break (DSB) mobility enhances the physical search for an ectopic template during homology-directed repair (HDR), we tested the effects of factors that control chromatin dynamics, including cohesin loading and kinetochore anchoring. The former but not the latter is altered in response to DSBs. Loss of the nonhistone high-mobility group protein Nhp6 reduces histone occupancy and increases chromatin movement, decompaction, and ectopic HDR. The loss of nucleosome remodeler INO80-C did the opposite, To see whether enhanced HDR depends on DSB mobility or the global chromatin response, we tested the ubiquitin ligase mutant uls1 Delta, which selectively impairs local but not global movement in response to a DSB. Strand invasion occurs in uls1 Delta cells with wild-type kinetics, arguing that global histone depletion rather than DSB movement is rate limiting for HDR. Impaired break movement in uls1 Delta correlates with elevated MRX and cohesin loading, despite normal resection and checkpoint activation.