Replication protein A promotes 5'-->3' end processing during homology-dependent DNA double-strand break repair.
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.
Fox Chase Cancer Center, Philadelphia, PA 19111, USA. Hong_Yan@fccc.edu
Hong
H
Yan
Thomas
T
Toczylowski
Jill
J
McCane
Chinyi
C
Chen
Shuren
S
Liao
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000378:metabolism; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D011485:Protein Binding; D051716:Replication Protein A / Q000378:metabolism*; D000071657:Werner Syndrome Helicase; D029867:Xenopus Proteins / Q000378:metabolism; D014982:Xenopus laevis
D004277:DNA, Single-Stranded; D051716:Replication Protein A; D029867:Xenopus Proteins; D004720:Endonucleases; D004265:DNA Helicases; C533186:DNA2 protein, Xenopus; D000071657:Werner Syndrome Helicase; C507159:WRN protein, Xenopus
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