Institute of Virology, University of Zurich, Zurich, Switzerland.
J Virol. 2012 Jan;86(1):143-55. doi: 10.1128/JVI.05694-11. Epub 2011 Oct 19.
Adeno-associated virus type 2 (AAV2) is a human parvovirus that relies on a helper virus for efficient replication. Herpes simplex virus 1 (HSV-1) supplies helper functions and changes the environment of the cell to promote AAV2 replication. In this study, we examined the accumulation of cellular replication and repair proteins at viral replication compartments (RCs) and the influence of replicating AAV2 on HSV-1-induced DNA damage responses (DDR). We observed that the ATM kinase was activated in cells coinfected with AAV2 and HSV-1. We also found that phosphorylated ATR kinase and its cofactor ATR-interacting protein were recruited into AAV2 RCs, but ATR signaling was not activated. DNA-PKcs, another main kinase in the DDR, was degraded during HSV-1 infection in an ICP0-dependent manner, and this degradation was markedly delayed during AAV2 coinfection. Furthermore, we detected phosphorylation of DNA-PKcs during AAV2 but not HSV-1 replication. The AAV2-mediated delay in DNA-PKcs degradation affected signaling through downstream substrates. Overall, our results demonstrate that coinfection with HSV-1 and AAV2 provokes a cellular DDR which is distinct from that induced by HSV-1 alone.
腺相关病毒 2 型（AAV2）是一种依赖辅助病毒进行有效复制的人类微小病毒。单纯疱疹病毒 1（HSV-1）提供辅助功能，并改变细胞环境以促进 AAV2 的复制。在这项研究中，我们研究了细胞复制和修复蛋白在病毒复制区（RC）的积累情况，以及复制 AAV2 对 HSV-1 诱导的 DNA 损伤反应（DDR）的影响。我们观察到在 AAV2 和 HSV-1 共感染的细胞中，ATM 激酶被激活。我们还发现磷酸化的 ATR 激酶及其辅助因子 ATR 相互作用蛋白被募集到 AAV2 RC 中，但 ATR 信号没有被激活。另一种 DDR 的主要激酶 DNA-PKcs 在 HSV-1 感染期间以 ICP0 依赖的方式被降解，而在 AAV2 共感染期间，这种降解明显延迟。此外，我们在 AAV2 复制期间检测到 DNA-PKcs 的磷酸化，但在 HSV-1 复制期间没有检测到。AAV2 介导的 DNA-PKcs 降解延迟影响了下游底物的信号转导。总的来说，我们的研究结果表明，HSV-1 和 AAV2 的共感染会引发不同于单纯 HSV-1 感染的细胞 DDR。
