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Mol Cell. 2013 May 23;50(4):589-600. doi: 10.1016/j.molcel.2013.04.032.

RPA coordinates DNA end resection and prevents formation of DNA hairpins.

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1
Department of Biological Sciences, Columbia University, New York, NY 10027, USA.

Abstract

Replication protein A (RPA) is an essential eukaryotic single-stranded DNA binding protein with a central role in DNA metabolism. RPA directly participates in DNA double-strand break repair by stimulating 5'-3' end resection by the Sgs1/BLM helicase and Dna2 endonuclease in vitro. Here we investigated the role of RPA in end resection in vivo, using a heat-inducible degron system that allows rapid conditional depletion of RPA in Saccharomyces cerevisiae. We found that RPA depletion eliminated both the Sgs1-Dna2- and Exo1-dependent extensive resection pathways and synergized with mre11Δ to prevent end resection. The short single-stranded DNA tails formed in the absence of RPA were unstable due to 3' strand loss and the formation of fold-back hairpin structures that required resection initiation and Pol32-dependent DNA synthesis. Thus, RPA is required to generate ssDNA, and also to protect ssDNA from degradation and inappropriate annealing that could lead to genome rearrangements.

PMID:
23706822
PMCID:
PMC3855855
DOI:
10.1016/j.molcel.2013.04.032
[Indexed for MEDLINE]
Free PMC Article

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