University of Missouri-Columbia, School of Medicine, Columbia, Missouri, United States of America.
PLoS Pathog. 2010 Oct 7;6(10):e1001141. doi: 10.1371/journal.ppat.1001141.
Infection by DNA viruses can elicit DNA damage responses (DDRs) in host cells. In some cases the DDR presents a block to viral replication that must be overcome, and in other cases the infecting agent exploits the DDR to facilitate replication. We find that low multiplicity infection with the autonomous parvovirus minute virus of mice (MVM) results in the activation of a DDR, characterized by the phosphorylation of H2AX, Nbs1, RPA32, Chk2 and p53. These proteins are recruited to MVM replication centers, where they co-localize with the main viral replication protein, NS1. The response is seen in both human and murine cell lines following infection with either the MVMp or MVMi strains. Replication of the virus is required for DNA damage signaling. Damage response proteins, including the ATM kinase, accumulate in viral-induced replication centers. Using mutant cell lines and specific kinase inhibitors, we show that ATM is the main transducer of the signaling events in the normal murine host. ATM inhibitors restrict MVM replication and ameliorate virus-induced cell cycle arrest, suggesting that DNA damage signaling facilitates virus replication, perhaps in part by promoting cell cycle arrest. Thus it appears that MVM exploits the cellular DNA damage response machinery early in infection to enhance its replication in host cells.
DNA 病毒感染会在宿主细胞中引发 DNA 损伤反应 (DDR)。在某些情况下,DDR 会对病毒复制造成阻碍,病毒必须克服这种阻碍;而在其他情况下,感染因子会利用 DDR 来促进复制。我们发现,低倍数感染自主细小病毒 minute virus of mice (MVM) 会导致 DDR 的激活,其特征是 H2AX、Nbs1、RPA32、Chk2 和 p53 的磷酸化。这些蛋白质被募集到 MVM 复制中心,与主要的病毒复制蛋白 NS1 共定位。在感染 MVMp 或 MVMi 株后,无论是在人源细胞系还是鼠源细胞系中都能观察到这种反应。病毒的复制是 DNA 损伤信号所必需的。包括 ATM 激酶在内的损伤反应蛋白在病毒诱导的复制中心积累。通过使用突变细胞系和特定的激酶抑制剂,我们表明 ATM 是正常鼠宿主中信号事件的主要转导器。ATM 抑制剂限制了 MVM 的复制并减轻了病毒诱导的细胞周期停滞,表明 DNA 损伤信号促进了病毒复制,这可能部分是通过促进细胞周期停滞来实现的。因此,MVM 似乎在感染早期利用细胞 DNA 损伤反应机制来增强其在宿主细胞中的复制。