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J Cell Biol January 24, 2011; 192 (2): 251-61.

Replication protein A promotes 5''-->3'' end processing during homology-dependent DNA double-strand break repair.

Yan H , Toczylowski T , McCane J , Chen C , Liao S .

Replication protein A (RPA), the eukaryotic single-strand deoxyribonucleic acid (DNA [ss-DNA])-binding protein, is involved in DNA replication, nucleotide damage repair, mismatch repair, and DNA damage checkpoint response, but its function in DNA double-strand break (DSB) repair is poorly understood. We investigated the function of RPA in homology-dependent DSB repair using Xenopus laevis nucleoplasmic extracts as a model system. We found that RPA is required for single-strand annealing, one of the homology-dependent DSB repair pathways. Furthermore, RPA promotes the generation of 3'' single-strand tails (ss-tails) by stimulating both the Xenopus Werner syndrome protein (xWRN)-mediated unwinding of DNA ends and the subsequent Xenopus DNA2 (xDNA2)-mediated degradation of the 5'' ss-tail. Purified xWRN, xDNA2, and RPA are sufficient to carry out the 5''-strand resection of DNA that carries a 3'' ss-tail. These results provide strong biochemical evidence to link RPA to a specific DSB repair pathway and reveal a novel function of RPA in the generation of 3'' ss-DNA for homology-dependent DSB repair.

PubMed ID: 21263027
PMC ID: PMC3172182
Article link: J Cell Biol
Grant support: [+]

Species referenced: Xenopus
Genes referenced: blm dna2 mre11 rbbp8 rpa1 rrad wrn

Article Images: [+] show captions
References [+] :
Baumann, Role of the human RAD51 protein in homologous recombination and double-stranded-break repair. 1998, Pubmed