title: RecA protein recruits structural maintenance of chromosomes (SMC)-like RecN protein to DNA double-strand breaks.
abstract: Escherichia coli RecN is an SMC (structural maintenance of chromosomes) family protein that is required for DNA double-strand break (DSB) repair. Previous studies show that GFP-RecN forms nucleoid-associated foci in response to DNA damage, but the mechanism by which RecN is recruited to the nucleoid is unknown. Here, we show that the assembly of GFP-RecN foci on the nucleoid in response to DNA damage involves a functional interaction between RecN and RecA. A novel RecA allele identified in this work, recA(Q300R), is proficient in SOS induction and repair of UV-induced DNA damage, but is deficient in repair of mitomycin C (MMC)-induced DNA damage. Cells carrying recA(Q300R) fail to recruit RecN to DSBs and accumulate fragmented chromosomes after exposure to MMC. The ATPase-deficient RecN(K35A) binds and forms foci at MMC-induced DSBs, but is not released from the MMC-induced DNA lesions, resulting in a defect in homologous recombination-dependent DSB repair. These data suggest that RecN plays a crucial role in homologous recombination-dependent DSB repair and that it is required upstream of RecA-mediated strand exchange.
authors: From the Department of Life Science, Graduate School of Science, Gakushuin University, Tokyo 171-8588.
authors: Kenji
authors: K
authors: Keyamura
authors: Chikako
authors: C
authors: Sakaguchi
authors: Yoshino
authors: Y
authors: Kubota
authors: Hironori
authors: H
authors: Niki
authors: Takashi
authors: T
authors: Hishida
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D053903:DNA Breaks, Double-Stranded; D004249:DNA Damage; D004262:DNA Restriction Enzymes / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016685:Mitomycin / Q000494:pharmacology; D020125:Mutation, Missense; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism*; D059767:Recombinational DNA Repair / Q000235:genetics; D013014:SOS Response, Genetics / Q000235:genetics
chemical_list: D000477:Alkylating Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D049452:Green Fluorescent Proteins; D016685:Mitomycin; D011938:Rec A Recombinases; D004262:DNA Restriction Enzymes; C047879:RecN protein, Bacteria
keywords: ATPases; DNA Damage; DNA Damage Response; DNA Double-strand Breaks; DNA Recombination; DNA Repair; DNA-Protein Interaction
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