RNF8 ubiquitinates RecQL4 and promotes its dissociation from DNA double strand breaks.
RNF8 ubiquitinates RecQL4 and promotes its dissociation from DNA double strand breaks.
复制标题
RNF8 泛素化 RecQL4 并促进其从 DNA 双链断裂中解离
DOI：
10.1038/s41389-021-00315-0
复制
发表时间：
2021-03-05
期刊：
影响因子：
6.2
通讯作者：
Zhao Y
中科院分区：
文献类型：
--
作者：
Tan Q;Niu K;Zhu Y;Chen Z;Li Y;Li M;Wei D;Balajee AS;Fang H;Zhao Y
泛素依赖的DNA损伤反应（DDR）信号在细胞选择DNA损伤修复途径中起着关键作用。人DNA解旋酶RecQL 4参与DNA复制和修复，RecQL 4的缺失与以基因组不稳定特征为特征的常染色体隐性遗传疾病相关。在早期的研究中，RecQL 4被分离为一种稳定的复合物，含有两种N端规则的泛素连接酶（UBR 1和UBR 2）。然而，目前还不清楚RecQL4泛素化状态是否对其DNA修复功能至关重要。在这里，我们报告说，RecQL 4直接与RNF8（环指泛素E3连接酶）相互作用，并在DNA双链断裂（DSB）位点共定位。我们的研究结果表明，RNF8泛素化RecQL4蛋白主要在876，1048和1101的赖氨酸位点，从而促进RecQL4从DSB位点的解离。RecQL4在泛素化位点的突变体在DSB上具有显著延长的保留，这阻碍了其直接下游DSB修复蛋白（CtIP和Ku80）的募集。有趣的是，在RecQL 4耗尽的细胞中观察到的降低的DSB修复能力仅通过野生型RecQL 4的重建而恢复，而不是泛素化突变体。此外，RecQL 4直接与WRAP 53 β相互作用，已知WRAP 53 β将RNF8募集到DSB，并且WRAP 53 β增强RecQL 4与RNF8的缔合。WRAP 53 β沉默导致RNF8向DSB的募集几乎减少，并且RecQL 4从DSB位点的解离大大减弱。总的来说，我们的研究表明，RNF8介导的泛素化事件构成了RecQL 4在DSB修复中功能的重要组成部分。
Ubiquitination-dependent DNA damage response (DDR) signals play a critical role in the cellular choice of DNA damage repair pathways. Human DNA helicase RecQL4 participates in DNA replication and repair, and loss of RecQL4 is associated with autosomal recessive genetic disorders characterized by genomic instability features. In an earlier study, RecQL4 was isolated as a stable complex that contained two ubiquitin ligases of the N-end rule (UBR1 and UBR2). However, it is unknown whether or not RecQL4 ubiquitination status is critical for its DNA repair function. Here, we report that RecQL4 directly interacts with RNF8 (a RING finger ubiquitin E3 ligase), and both co-localize at DNA double-strand break (DSB) sites. Our findings indicate that RNF8 ubiquitinates RecQL4 protein mainly at the lysine sites of 876, 1048, and 1101, thereby facilitating the dissociation of RecQL4 from DSB sites. RecQL4 mutant at ubiquitination sites had a significantly prolonged retention at DSBs, which hinders the recruitment of its direct downstream DSB repair proteins (CtIP & Ku80). Interestingly, reduced DSB repair capacity observed in RecQL4 depleted cells was restored only by the reconstitution of wild-type RecQL4, but not the ubiquitination mutant. Additionally, RecQL4 directly interacts with WRAP53β that is known to recruit RNF8 to DSBs and WRAP53β enhances the association of RecQL4 with RNF8. WRAP53β silencing resulted in a nearly diminished recruitment of RNF8 to DSBs and in a greatly attenuated dissociation of RecQL4 from the DSB sites. Collectively, our study demonstrates that the ubiquitination event mediated by RNF8 constitutes an essential component for RecQL4’s function in DSB repair.
登录
查看更多内容
DOI：
10.18632/aging.100334
发表时间：
2011-05
期刊：
Aging
影响因子：
--
作者：
Harris JL;Khanna KK
通讯作者：
Khanna KK
影响因子：
10.5
作者：
Henriksson S;Rassoolzadeh H;Hedström E;Coucoravas C;Julner A;Goldstein M;Imreh G;Zhivotovsky B;Kastan MB;Helleday T;Farnebo M
通讯作者：
Farnebo M
RecQL4-Aurora B 激酶轴对于细胞增殖、细胞周期进展和有丝分裂完整性至关重要。
影响因子：
6.2
作者：
Fang H;Niu K;Mo D;Zhu Y;Tan Q;Wei D;Li Y;Chen Z;Yang S;Balajee AS;Zhao Y
通讯作者：
Zhao Y
影响因子：
16
作者：
Paul A;Wang B
通讯作者：
Wang B
影响因子：
14.9
作者：
Pandita TK;Richardson C
通讯作者：
Richardson C