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首页> 外文期刊>Molecular and Cellular Biology >Conservative Repair of a Chromosomal Double-Strand Break by Single-Strand DNA through Two Steps of Annealing
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Conservative Repair of a Chromosomal Double-Strand Break by Single-Strand DNA through Two Steps of Annealing

机译：通过两步退火，单链DNA保守修复染色体双链断裂

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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.
机译：染色体双链断裂（DSB）的修复对于正常细胞的生长至关重要，而同源重组是DSB修复的普遍过程。我们使用与DSB两侧具有同源性的单链寡核苷酸探索了酿酒酵母（Saccharomyces cerevisiae）中的DSB修复机制。寡核苷酸定向修复仅通过Rad52和Rad59介导的单链退火（SSA）进行。甚至人Rad52的SSA域也为 rad52 无效突变提供了部分互补。修复不涉及Rad51驱动的链入侵，而且链入侵的抑制增加了寡核苷酸的修复。显示DSB可通过与链断裂偏距较大距离（至少20 kb）的断裂端一侧同源的寡核苷酸以链偏向的方式激活靶向，表明广泛的5'至3'切除，随后恢复将DNA切割成双链状态。我们得出的结论是，通过两轮退火，单链DNA寡核苷酸可修复长久切除的染色体DSB末端。单链DNA的修复可能是保守的，并且可能允许使用间隔较近的DSB准确还原染色体DNA。

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