In vivo roles of Rad52, Rad54, and Rad55 proteins in Rad51-mediated recombination
In vivo roles of Rad52, Rad54, and Rad55 proteins in Rad51-mediated recombination
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DOI：
10.1016/s1097-2765(03)00269-7
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发表时间：
2003-07-01
期刊：
影响因子：
16
通讯作者：
Haber, JE
中科院分区：
文献类型：
--
作者：
Sugawara, N;Wang, X;Haber, JE
通过同源重组修复双链断裂需要将链交换蛋白Rad 51 p结合到ssDNA，然后与同源供体突触。在这里，我们使用染色质免疫沉淀监测在体内协会的酿酒酵母Rad 51 p与切割的MATa基因座和HMLalpha供体。Rad 51 p定位于MAT之前，它与HML的协会，提供了证据的时间需要为Rad 51丝搜索基因组的同源序列。Rad 51 p与ssDNA的结合需要Rad 52 p。Rad 55 p的缺失延迟了Rad 51 p与ssDNA的结合，并阻止了链侵入和Rad 51 p定位于HMLalpha。缺乏Rad 54 p不会显著损害Rad 51 p向MAT的募集或其与HMLalpha的初始结合;然而，在侵入链的3'端发生突触后，在DNA合成起始时或之前需要Rad 54 p。
Repairing a double-strand break by homologous recombination requires binding of the strand exchange protein Rad51p to ssDNA, followed by synapsis with a homologous donor. Here we used chromatin immunoprecipitation to monitor the in vivo association of Saccharomyces cerevisiae Rad51p with both the cleaved MATa locus and the HMLalpha donor. Localization of Rad51p to MAT precedes its association with HML, providing evidence of the time needed for the Rad51 filament to search the genome for a homologous sequence. Rad51p binding to ssDNA requires Rad52p. The absence of Rad55p delays Rad51p binding to ssDNA and prevents strand invasion and localization of Rad51p to HMLalpha. Lack of Rad54p does not significantly impair Rad51p recruitment to MAT or its initial association with HMLalpha; however, Rad54p is required at or before the initiation of DNA synthesis after synapsis has occurred at the 3' end of the invading strand.