REV1是跨损伤聚合酶Y家族的重要成员之一,它不仅作为支架蛋白介导Y家族聚合酶招募至损伤位点完成跨损伤DNA合成(translesion DNA synthesis, TLS),还可利用自身的dCMP转移酶活性在一些损伤位点对侧整合dCMP参与TLS。此外,REV1也被报导...REV1是跨损伤聚合酶Y家族的重要成员之一,它不仅作为支架蛋白介导Y家族聚合酶招募至损伤位点完成跨损伤DNA合成(translesion DNA synthesis, TLS),还可利用自身的dCMP转移酶活性在一些损伤位点对侧整合dCMP参与TLS。此外,REV1也被报导参与调控同源重组修复。为进一步探讨REV1互作蛋白RAD51和RAD51C在其参与的同源重组修复通路中的调控作用,本研究采用脉冲氮激光微辐射实验,发现RAD51可调控REV1到双链断裂位点的募集。同时,免疫荧光实验结果证明REV1也反过来影响RAD51应答CPT损伤。然而敲低RAD51C并不影响REV1到DNA双链断裂位点的招募。结果表明,REV1和RAD51在HR通路中存在彼此相互调控的关系。展开更多
In addition to DNA repair pathways,cells utilize translesion DNA synthesis(TLS)to bypass DNA lesions during replication.During TLS,Y-family DNA polymerase(Polη,Polκ,Polιand Rev1)inserts specific nucleotide opposite...In addition to DNA repair pathways,cells utilize translesion DNA synthesis(TLS)to bypass DNA lesions during replication.During TLS,Y-family DNA polymerase(Polη,Polκ,Polιand Rev1)inserts specific nucleotide opposite preferred DNA lesions,and then Polζ consisting of two subunits,Rev3 and Rev7,carries out primer extension.Here,we report the complex structures of Rev3-Rev7-Rev1^(CTD) and Rev3-Rev7-Rev1^(CTD)-Polκ^(RIR).These two structures demonstrate that Rev1^(CTD) contains separate binding sites for Polκand Rev7.Our BIAcore experiments provide additional support for the notion that the interaction between Rev3 and Rev7 increases the affinity of Rev7 and Rev1.We also verified through FRET experiment that Rev1,Rev3,Rev7 and Polκ form a stable quaternary complex in vivo,thereby suggesting an efficient switching mechanism where the“inserter”polymerase can be immediately replaced by an“extender”polymerase within the same quaternary complex.展开更多
文摘REV1是跨损伤聚合酶Y家族的重要成员之一,它不仅作为支架蛋白介导Y家族聚合酶招募至损伤位点完成跨损伤DNA合成(translesion DNA synthesis, TLS),还可利用自身的dCMP转移酶活性在一些损伤位点对侧整合dCMP参与TLS。此外,REV1也被报导参与调控同源重组修复。为进一步探讨REV1互作蛋白RAD51和RAD51C在其参与的同源重组修复通路中的调控作用,本研究采用脉冲氮激光微辐射实验,发现RAD51可调控REV1到双链断裂位点的募集。同时,免疫荧光实验结果证明REV1也反过来影响RAD51应答CPT损伤。然而敲低RAD51C并不影响REV1到DNA双链断裂位点的招募。结果表明,REV1和RAD51在HR通路中存在彼此相互调控的关系。
文摘基于核级控制阀的安全性和经济性的考虑,首先介绍了核级部件设计制造过程的等级划分,重点介绍了核级控制阀设计制造过程中的商品级部件升级方法(Commercial Grade Dedication Method)的依据标准EPRI NP-5652&TR-102260 Rev.1①(Plant Engineering:Guideline for the acceptance of Commercial-Grade Items In Nuclear Safety-Related Applications),并对商品级部件升级的验收方法进行了讨论。
基金supported financially by the National Basic Research Program(973 Program)(No.2011CB910302)the National Natural Science Foundation of China(Grant Nos.31025009,31021062 and 31200558).
文摘In addition to DNA repair pathways,cells utilize translesion DNA synthesis(TLS)to bypass DNA lesions during replication.During TLS,Y-family DNA polymerase(Polη,Polκ,Polιand Rev1)inserts specific nucleotide opposite preferred DNA lesions,and then Polζ consisting of two subunits,Rev3 and Rev7,carries out primer extension.Here,we report the complex structures of Rev3-Rev7-Rev1^(CTD) and Rev3-Rev7-Rev1^(CTD)-Polκ^(RIR).These two structures demonstrate that Rev1^(CTD) contains separate binding sites for Polκand Rev7.Our BIAcore experiments provide additional support for the notion that the interaction between Rev3 and Rev7 increases the affinity of Rev7 and Rev1.We also verified through FRET experiment that Rev1,Rev3,Rev7 and Polκ form a stable quaternary complex in vivo,thereby suggesting an efficient switching mechanism where the“inserter”polymerase can be immediately replaced by an“extender”polymerase within the same quaternary complex.
