Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM).
Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM).
复制标题
小鼠微小病毒 (MVM) NS2 突变体感染期间，DNA 复制和损伤反应蛋白募集到病毒复制中心。
DOI：
10.1016/j.virol.2010.12.009
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Tattersall,Peter
中科院分区：
文献类型：
--
作者：
Ruiz,Zandra;Mihaylov,IvailoS;Cotmore,SusanF;Tattersall,Peter
MVM NS 2是病毒DNA扩增所必需的，但其作用机制尚不清楚。自主细小病毒相关复制（APAR）中心发展的分类方案，基于NS 1分布，被用来表征异常APAR体成熟NS 2无效突变体感染，和他们的组织检查宿主蛋白募集的缺陷。由于获得已知的复制因子似乎是正常的，我们寻找调用DNA损伤反应的差异。我们观察到H2 AX/MDC 1损伤反应灶与病毒复制中心的广泛关联，以及发生在野生型和突变感染中的RPA 32的隔离和复杂的过度磷酸化。通过Western转移定量这些反应表明，野生型和NS 2突变体MVM都引起ATM激活，而在异步A9细胞中已经基本激活的ATR的磷酸化被下调。我们的结论是，MVM感染引起多种损伤反应，影响APAR环境，但NS 2不修改细胞蛋白的招聘。
MVM NS2 is essential for viral DNA amplification, but its mechanism of action is unknown. A classification scheme for autonomous parvovirus-associated replication (APAR) center development, based on NS1 distribution, was used to characterize abnormal APAR body maturation in NS2null mutant infections, and their organization examined for defects in host protein recruitment. Since acquisition of known replication factors appeared normal, we looked for differences in invoked DNA damage responses. We observed widespread association of H2AX/MDC1 damage response foci with viral replication centers, and sequestration and complex hyperphosphorylation of RPA32, which occurred in wildtype and mutant infections. Quantifying these responses by western transfer indicated that both wildtype and NS2 mutant MVM elicited ATM activation, while phosphorylation of ATR, already basally activated in asynchronous A9 cells, was downregulated. We conclude that MVM infection invokes multiple damage responses that influence the APAR environment, but that NS2 does not modify the recruitment of cellular proteins.