Recruitment of the recombinational repair machinery to a DNA double-strand break in yeast
Recruitment of the recombinational repair machinery to a DNA double-strand break in yeast
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DOI：
10.1016/s1097-2765(03)00242-9
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发表时间：
2003-07-01
期刊：
影响因子：
16
通讯作者：
Peterson, CL
中科院分区：
文献类型：
--
作者：
Wolner, B;van Komen, S;Peterson, CL
通过同源重组修复 DNA 双链断裂 (DSB) 需要 RAD52 上位组的成员。在这里，我们使用染色质免疫沉淀 (ChIP) 来检查 Rad51p、Rad52p、Rad54p、Rad55p 和 RPA 在酵母中招募到单个诱导 DSB 的时间顺序。我们的结果表明，通过 Rad51p 的结合启动邻近 DSB 的 Rad 蛋白的连续、相互依赖的组装。各种突变株的 ChIP 时间进程和其他生化研究表明 Rad52p、Rad55p 和 Rad54p 各自有助于促进 Rad51p 核蛋白丝的形成和/或稳定。我们还发现，所有四种 Rad 蛋白在链入侵过程中都与同源供体序列相关。这些研究提供了功能性 Rad51p 突触前丝体内组装所需分子事件的近乎全面的了解。
Repair of DNA double-strand breaks (DSBs) by homologous recombination requires members of the RAD52 epistasis group. Here we use chromatin immunoprecipitation (ChIP) to examine the temporal order of recruitment of Rad51p, Rad52p, Rad54p, Rad55p, and RPA to a single, induced DSB in yeast. Our results suggest a sequential, interdependent assembly of Rad proteins adjacent to the DSB initiated by binding of Rad51p. ChIP time courses from various mutant strains and additional biochemical studies suggest that Rad52p, Rad55p, and Rad54p each help promote the formation and/or stabilization of the Rad51p nucleoprotein filament. We also find that all four Rad proteins associate with homologous donor sequences during strand invasion. These studies provide a near comprehensive view of the molecular events required for the in vivo assembly of a functional Rad51p presynaptic filament.