Conservative repair of a chromosomal double-strand break by single-strand DNA through two steps of annealing

被引:81
作者
Storici, Francesca [1 ]
Snipe, Joyce R. [1 ]
Chan, Godwin K. [1 ]
Gordenin, Dmitry A. [1 ]
Resnick, Michael A. [1 ]
机构
[1] NIEHS, Genet Mol Lab, Head Chromosome Stabil Sect, NIH, Res Triangle Pk, NC 27709 USA
关键词
D O I
10.1128/MCB.00672-06
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The repair of chromosomal double-strand breaks (DSBs) is essential to normal cell growth, and homologous recombination is a universal process for DSB repair. We explored DSB repair mechanisms in the yeast Saccharomyces cerevisiae using single-strand oligonucleotides with homology to both sides of a DSB. Oligonucleotide-directed repair occurred exclusively via Rad52- and Rad59-mediated single-strand annealing (SSA). Even the SSA domain of human Rad52 provided partial complementation for a null rad52 mutation. The repair did not involve Rad51-driven strand invasion, and moreover the suppression of strand invasion increased repair with oligonucleotides. A DSB was shown to activate targeting by oligonucleotides homologous to only one side of the break at large distances (at least 20 kb) from the break in a strand-biased manner, suggesting extensive 5' to 3' resection, followed by the restoration of resected DNA to the double-strand state. We conclude that long resected chromosomal DSB ends are repaired by a single-strand DNA oligonucleotide through two rounds of annealing. The repair by single-strand DNA can be conservative and may allow for accurate restoration of chromosomal DNAs with closely spaced DSBs.
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页码:7645 / 7657
页数:13
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