Wilkinson Dianna E, Weller Sandra K
Department of Molecular, Microbial and Structural Biology MC3205, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
J Cell Sci. 2006 Jul 1;119(Pt 13):2695-703. doi: 10.1242/jcs.02981. Epub 2006 Jun 6.
Like other DNA viruses, herpes simplex virus type 1 (HSV-1) interacts with components of the cellular response to DNA damage. For example, HSV-1 sequesters endogenous, uninduced, hyperphosphorylated RPA (replication protein A) away from viral replication compartments. RPA is a ssDNA-binding protein that signals genotoxic stress through the ATR (ataxia telangiectasia-mutated and Rad3-related) pathway. The sequestration of endogenous hyperphosphorylated RPA away from replicating viral DNA suggests that HSV-1 prevents the normal ATR-signaling response. In this study we examine the spatial distribution of endogenous hyperphosphorylated RPA with respect to ATR, its recruitment factor, ATRIP, and the cellular dsDNA break marker, gammaH2AX, during HSV-1 infection. The accumulation of these repair factors at DNA lesions has previously been identified as an early event in signaling genotoxic stress. We show that HSV-1 infection disrupts the ATR pathway by a mechanism that prevents the recruitment of repair factors, spatially uncouples ATRIP from ATR and sequesters ATRIP and endogenous hyperphosphorylated RPA within virus-induced nuclear domains containing molecular chaperones and components of the ubiquitin proteasome. The HSV-1 immediate early protein ICP0 is sufficient to induce the redistribution of ATRIP. This is the first report that a virus can disrupt the usually tight colocalization of ATR and ATRIP.
与其他DNA病毒一样,单纯疱疹病毒1型(HSV-1)与细胞对DNA损伤的反应成分相互作用。例如,HSV-1将内源性、未诱导的、高度磷酸化的复制蛋白A(RPA)从病毒复制区隔中隔离出来。RPA是一种单链DNA结合蛋白,通过共济失调毛细血管扩张突变和Rad3相关蛋白(ATR)途径发出基因毒性应激信号。将内源性高度磷酸化的RPA从复制的病毒DNA中隔离出来表明,HSV-1可阻止正常的ATR信号反应。在本研究中,我们检测了HSV-1感染期间内源性高度磷酸化RPA相对于ATR、其募集因子ATR相互作用蛋白(ATRIP)以及细胞双链DNA断裂标志物γH2AX的空间分布。这些修复因子在DNA损伤处的积累先前已被确定为发出基因毒性应激信号的早期事件。我们发现,HSV-1感染通过一种机制破坏ATR途径,该机制可阻止修复因子的募集,使ATRIP在空间上与ATR分离,并将ATRIP和内源性高度磷酸化的RPA隔离在含有分子伴侣和泛素蛋白酶体成分的病毒诱导核域内。HSV-1的立即早期蛋白ICP0足以诱导ATRIP的重新分布。这是关于病毒可破坏ATR和ATRIP通常紧密共定位的首次报道。