rad50 is required for efficient initiation of resection and recombinational repair at random, γ-induced double-strand break endsrad50需要有效的起始切除和随机重组修复,γ-induced双链断裂.pdfVIP

RAD50 Is Required for Efficient Initiation of Resection and Recombinational Repair at Random, c-Induced Double-Strand Break Ends 1 1 1 2 2 1 Jim Westmoreland , Wenjian Ma , Yan Yan , Kelly Van Hulle , Anna Malkova , Michael A. Resnick * 1 Chromosome Stability Section, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, United States of America, 2 Biology Department, Indiana University–Purdue University, Indianapolis, Indiana, United States of America Abstract Resection of DNA double-strand break (DSB) ends is generally considered a critical determinant in pathways of DSB repair and genome stability. Unlike for enzymatically induced site-specific DSBs, little is known about processing of random ‘‘dirty- ended’’ DSBs created by DNA damaging agents such as ionizing radiation. Here we present a novel system for monitoring early events in the repair of random DSBs, based on our finding that single-strand tails generated by resection at the ends of large molecules in budding yeast decreases mobility during pulsed field gel electrophoresis (PFGE). We utilized this ‘‘PFGE- shift’’ to follow the fate of both ends of linear molecules generated by a single random DSB in circular chromosomes. Within 10 min after c-irradiation of G2/M arrested WT cells, there is a near-synchronous PFGE-shift of the linearized circular molecules, corresponding to resection of a few hundred bases. Resection at the radiation-induced DSBs continues so that by the time of significant repair of DSBs at 1 hr there is about 1–2 kb resection per DSB end. The PFGE-shift is comparable in WT and recombination-defective rad52 and rad51 strains but somewhat delayed in exo1 mutants. However, in rad50 an