Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan.
Nucleic Acids Res. 2010 Aug;38(15):5059-74. doi: 10.1093/nar/gkq271. Epub 2010 Apr 19.
RAD51 is a key factor in homologous recombination (HR) and plays an essential role in cellular proliferation by repairing DNA damage during replication. The assembly of RAD51 at DNA damage is strictly controlled by RAD51 mediators, including BRCA1 and BRCA2. We found that human RAD51 directly binds GEMIN2/SIP1, a protein involved in spliceosome biogenesis. Biochemical analyses indicated that GEMIN2 enhances the RAD51-DNA complex formation by inhibiting RAD51 dissociation from DNA, and thereby stimulates RAD51-mediated homologous pairing. GEMIN2 also enhanced the RAD51-mediated strand exchange, when RPA was pre-bound to ssDNA before the addition of RAD51. To analyze the function of GEMIN2, we depleted GEMIN2 in the chicken DT40 line and in human cells. The loss of GEMIN2 reduced HR efficiency and resulted in a significant decrease in the number of RAD51 subnuclear foci, as observed in cells deficient in BRCA1 and BRCA2. These observations and our biochemical analyses reveal that GEMIN2 regulates HR as a novel RAD51 mediator.
RAD51 是同源重组(HR)的关键因素,在复制过程中修复 DNA 损伤,对细胞增殖起着至关重要的作用。RAD51 在 DNA 损伤处的组装受到 RAD51 介体的严格控制,包括 BRCA1 和 BRCA2。我们发现人类 RAD51 可直接与 GEMIN2/SIP1 结合,后者是参与剪接体生物发生的蛋白质。生化分析表明,GEMIN2 通过抑制 RAD51 从 DNA 上解离来增强 RAD51-DNA 复合物的形成,从而刺激 RAD51 介导的同源配对。当 RAD51 加入之前 RPA 已预结合 ssDNA 时,GEMIN2 还增强了 RAD51 介导的链交换。为了分析 GEMIN2 的功能,我们在鸡 DT40 系和人细胞中耗尽 GEMIN2。GEMIN2 的缺失降低了 HR 效率,并导致 RAD51 亚核焦点数量显著减少,这在 BRCA1 和 BRCA2 缺陷细胞中观察到。这些观察结果和我们的生化分析表明,GEMIN2 作为一种新的 RAD51 介体调节 HR。