N-terminal region of RecQ4 inhibits non-homologous end joining and chromatin association of the Ku heterodimer in Xenopus egg extracts
N-terminal region of RecQ4 inhibits non-homologous end joining and chromatin association of the Ku heterodimer in Xenopus egg extracts
复制标题
RecQ4 的 N 末端区域抑制非洲爪蟾卵提取物中 Ku 异二聚体的非同源末端连接和染色质关联
DOI：
10.1016/j.gene.2021.145647
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发表时间：
2021
期刊：
影响因子：
3.5
通讯作者：
Tada Shusuke
中科院分区：
文献类型：
--
作者：
Tsuyama Takashi;Fujita Kumiko;Sasaki Ryosuke;Hamanaka Shiori;Sotoyama Yuki;Ogawa Akira;Kusuzaki Kana;Azuma Yutaro;Tada Shusuke
RecQ 4是RecQ解旋酶家族的成员，是维持基因组完整性所必需的。RecQ 4已经显示出促进以下两种DNA双链断裂（DSB）修复途径：非同源末端连接（NHEJ）和同源重组（HR）。然而，其分子功能尚未完全阐明。在本研究中，我们的目的是研究RecQ 4在NHEJ使用Xenopusegg提取物的作用。将XenopusRecQ 4（N598）的N-末端598个氨基酸区域（其缺少中心解旋酶结构域和下游C-末端区域）添加到提取物中，并分析其对DNA末端连接的影响。我们发现N598抑制提取物中线性化DNA末端的连接。此外，N598抑制DSB诱导的Ku 70的染色质结合，这是NHEJ所必需的，而DSB诱导的HR相关蛋白，复制蛋白A（RPA）和Rad 51的染色质结合增加后，N598的加入。这些结果表明RecQ 4可能通过影响Ku异源二聚体与DNA末端的结合来影响DSB修复途径的选择。
RecQ4, a member of the RecQ helicase family, is required for the maintenance of genome integrity. RecQ4 has been shown to promote the following two DNA double-strand break (DSB) repair pathways: non-homologous end joining (NHEJ) and homologous recombination (HR). However, its molecular function has not been fully elucidated. In the present study, we aimed to investigate the role of RecQ4 in NHEJ usingXenopusegg extracts. The N-terminal 598 amino acid region ofXenopusRecQ4 (N598), which lacks a central helicase domain and a downstream C-terminal region, was added to the extracts and its effect on the joining of DNA ends was analyzed. We found that N598 inhibited the joining of linearized DNA ends in the extracts. In addition, N598 inhibited DSB-induced chromatin binding of Ku70, which is essential for NHEJ, while the DSB-induced chromatin binding of the HR-associated proteins, replication protein A (RPA) and Rad51, increased upon the addition of N598. These results suggest that RecQ4 possibly influences the choice of the DSB repair pathway by influencing the association of the Ku heterodimer with the DNA ends.