Recruitment of DNA repair synthesis machinery to sites of DNA damage/repair in living human cells.
Recruitment of DNA repair synthesis machinery to sites of DNA damage/repair in living human cells.
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DOI：
10.1093/nar/gkm115
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发表时间：
2007
影响因子：
14.9
通讯作者：
Yasui A
中科院分区：
文献类型：
--
作者：
Hashiguchi K;Matsumoto Y;Yasui A
真核细胞的DNA滑动钳，即增殖细胞核抗原，是DNA复制和修复合成所必需的。为了将环状、同源三聚体的增殖细胞核抗原加载到DNA双螺旋上，需要复制因子C(RFC)钳制加载器复合体的ATPase活性。虽然RFC向DNA复制位点募集了增殖细胞核抗原，但我们对其在DNA修复合成过程中募集的了解是有限的。在这项研究中，我们分析了在UVA-激光微照射造成HeLa细胞DNA损伤的局部部位，DNA修复合成所需的内源性和荧光标记蛋白的积累。RFC大亚基(RFC140)的聚集动力学和体外下拉实验表明，RFC对DNA损伤有两种不同的募集模式，一种是由于增殖细胞核抗原与RFC140的N端相互作用而导致的RFC140和增殖细胞核抗原的同时聚集，另一种是RFC140在DNA损伤部位的聚集比增殖细胞核抗原快得多。此外，RFC140基因敲除实验表明，在DNA损伤时，增殖细胞核抗原可以独立于RFC而积聚。这些结果表明，DNA损伤时RFC和增殖细胞核抗原的即刻积累只是部分相互依赖的。
The eukaryotic sliding DNA clamp, proliferating cell nuclear antigen (PCNA), is essential for DNA replication and repair synthesis. In order to load the ring-shaped, homotrimeric PCNA onto the DNA double helix, the ATPase activity of the replication factor C (RFC) clamp loader complex is required. Although the recruitment of PCNA by RFC to DNA replication sites has well been documented, our understanding of its recruitment during DNA repair synthesis is limited. In this study, we analyzed the accumulation of endogenous and fluorescent-tagged proteins for DNA repair synthesis at the sites of DNA damage produced locally by UVA-laser micro-irradiation in HeLa cells. Accumulation kinetics and in vitro pull-down assays of the large subunit of RFC (RFC140) revealed that there are two distinct modes of recruitment of RFC to DNA damage, a simultaneous accumulation of RFC140 and PCNA caused by interaction between PCNA and the extreme N-terminus of RFC140 and a much faster accumulation of RFC140 than PCNA at the damaged site. Furthermore, RFC140 knock-down experiments showed that PCNA can accumulate at DNA damage independently of RFC. These results suggest that immediate accumulation of RFC and PCNA at DNA damage is only partly interdependent.
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影响因子：
10.5
作者：
Zou, L;Cortez, D;Elledge, SJ
通讯作者：
Elledge, SJ
影响因子：
14.9
作者：
Fan, JS;Otterlei, M;Wilson, DM
通讯作者：
Wilson, DM
DOI：
10.1016/j.bbrc.2006.07.037
发表时间：
2006-09-15
影响因子：
3.1
作者：
Karmakar, Parimal;Seki, Masayuki;Enomoto, Takemi
通讯作者：
Enomoto, Takemi
影响因子：
5.3
作者：
Okano, S;Lan, L;Yasui, R
通讯作者：
Yasui, R
DOI：
10.1073/pnas.0406048101
发表时间：
2004-09-21
影响因子：
11.1
作者：
Lan, L;Nakajima, S;Yasui, A
通讯作者：
Yasui, A