XRCC1 is phosphorylated by DNA-dependent protein kinase in response to DNA damage.
XRCC1 is phosphorylated by DNA-dependent protein kinase in response to DNA damage.
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DOI：
10.1093/nar/gkj409
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发表时间：
2006
影响因子：
14.9
通讯作者：
Ménissier-de Murcia J
中科院分区：
文献类型：
--
作者：
Lévy N;Martz A;Bresson A;Spenlehauer C;de Murcia G;Ménissier-de Murcia J
XRCC1 的两个 BRCT 结构域（BRCT1 和 BRCT2）通过 DNA 单链断裂 (SSB) 和碱基损伤修复途径的几个关键因素介导蛋白质-蛋白质相互作用网络。 BRCT1 是 XRCC1 立即依赖多聚（ADP-核糖）募集到 DNA 断裂处所必需的，并且对于 DNA 损伤后的生存至关重要。为了更好地了解 XRCC1 在 DNA 末端加工中的生物学作用，通过对 HeLa 细胞提取物中的 GST-BRCT1 下拉蛋白进行质谱分析来寻找 BRCT1 结构域相关蛋白。在这里，我们报告双链断裂（DSB）修复异三聚体复合物DNA-PK与XRCC1的BRCT1结构域相互作用，并在电离辐射后磷酸化该结构域的丝氨酸371。这导致 XRCC1 二聚体解离。 XRCC1 R399Q 变异等位基因不影响这种磷酸化。我们还表明 XRCC1 通过 DNA-PK 强烈刺激 p53-Ser15 的磷酸化。假磷酸化S371D突变体是DNA-PK活性较弱的刺激剂，而具有DNA-PK刺激能力的非磷酸化突变体S371L未能完全挽救XRCC1缺陷的EM9啮齿动物细胞的DSB修复缺陷。 XRCC1 和 DNA-PK 之间响应 IR 的功能关联为它们参与共同的 DSB 修复途径提供了第一个证据。
The two BRCT domains (BRCT1 and BRCT2) of XRCC1 mediate a network of protein–protein interactions with several key factors of the DNA single-strand breaks (SSBs) and base damage repair pathways. BRCT1 is required for the immediate poly(ADP–ribose)-dependent recruitment of XRCC1 to DNA breaks and is essential for survival after DNA damage. To better understand the biological role of XRCC1 in the processing of DNA ends, a search for the BRCT1 domain-associated proteins was performed by mass spectrometry of GST-BRCT1 pulled-down proteins from HeLa cell extracts. Here, we report that the double-strand break (DSB) repair heterotrimeric complex DNA-PK interacts with the BRCT1 domain of XRCC1 and phosphorylates this domain at serine 371 after ionizing irradiation. This caused XRCC1 dimer dissociation. The XRCC1 R399Q variant allele did not affect this phosphorylation. We also show that XRCC1 strongly stimulates the phosphorylation of p53-Ser15 by DNA-PK. The pseudo phosphorylated S371D mutant was a much weaker stimulator of DNA-PK activity whereas the non-phosphorylable mutant S371L endowed with a DNA-PK stimulating capacity failed to fully rescue the DSB repair defect of XRCC1-deficient EM9 rodent cells. The functional association between XRCC1 and DNA-PK in response to IR provides the first evidence for their involvement in a common DSB repair pathway.
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影响因子：
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