Herpes simplex virus 1 ICP0 phosphorylation site mutants are attenuated for viral replication and impaired for explant-induced reactivation.
Herpes simplex virus 1 ICP0 phosphorylation site mutants are attenuated for viral replication and impaired for explant-induced reactivation.
复制标题
单纯疱疹病毒 1 ICP0 磷酸化位点突变体的病毒复制减弱，外植体诱导的再激活受损。
DOI：
10.1128/jvi.05661-11
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发表时间：
2011
影响因子：
5.4
通讯作者：
Davido,DavidJ
中科院分区：
文献类型：
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作者：
Mostafa,HebaH;Thompson,ThorntonW;Kushnir,AnnaS;Haenchen,SteveD;Bayless,AdamM;Hilliard,JoshuaG;Link,MalenA;Pitcher,LisaA;Loveday,Emma;Schaffer,PriscillaA;Davido,DavidJ
在细胞培养实验中，磷酸化似乎是单纯疱疹病毒1（HSV-1）即早期（IE）蛋白ICP 0的关键调节因子，ICP 0是一种反式激活病毒基因表达的E3泛素连接酶。已经鉴定了ICP 0中磷酸化的三个主要区域（氨基酸224至232、365至371和508至518），并且已经构建了阻断每个区域内磷酸化位点的突变病毒（分别称为Phos 1、2和3）。先前的研究表明，与野生型病毒相比，Phos 1在细胞培养物中的复制显著减少（C. Boutell等人，J. Virol. 82：10647-10656，2008）。为了确定这些磷酸化位点突变对病毒体内生命周期的影响，用野生型HSV-1、Phos突变体或它们的标记拯救对应物对小鼠进行眼部感染。随后，检查了这些病毒的病毒复制、潜伏期的建立和病毒灭活诱导的再活化。相对于野生型病毒，Phos 1眼滴度在感染后第1天和第5天分别降低了7倍和18倍。在感染后第1天，Phos 2眼滴度显示降低6倍。在感染后第5天，Phos 1和2三叉神经节的滴度分别降低了16倍和20倍。此外，Phos 1和2的再活化效率相对于野生型HSV-1受损，尽管两种病毒在体内建立了野生型潜伏水平。Phos 3的急性复制、潜伏期和再活化表型与野生型HSV-1相似。我们从这些研究中得出结论，在HSV-1感染的小鼠眼部模型中，磷酸化可能是ICP 0生物活性的关键调节剂。
In cell culture experiments, phosphorylation appears to be a critical regulator of the herpes simplex virus 1 (HSV-1) immediate-early (IE) protein, ICP0, which is an E3 ubiquitin ligase that transactivates viral gene expression. Three major regions of phosphorylation in ICP0 (amino acids 224 to 232, 365 to 371, and 508 to 518) have been identified, and mutant viruses that block phosphorylation sites within each region (termed Phos 1, 2, and 3, respectively) have been constructed. Previous studies indicated that replication of Phos 1 is significantly reduced compared to that of wild-type virus in cell culture (C. Boutell, et al., J. Virol. 82:10647–10656, 2008). To determine the effects these phosphorylation site mutations have on the viral life cyclein vivo, mice were ocularly infected with wild-type HSV-1, the Phos mutants, or their marker rescue counterparts. Subsequently, viral replication, establishment of latency, and viral explant-induced reactivation of these viruses were examined. Relative to wild-type virus, Phos 1 eye titers were reduced as much as 7- and 18-fold on days 1 and 5 postinfection, respectively. Phos 2 eye titers showed a decrease of 6-fold on day 1 postinfection. Titers of Phos 1 and 2 trigeminal ganglia were reduced as much as 16- and 20-fold, respectively, on day 5 postinfection. Additionally, the reactivation efficiencies of Phos 1 and 2 were impaired relative to wild-type HSV-1, although both viruses established wild-type levels of latencyin vivo. The acute replication, latency, and reactivation phenotypes of Phos 3 were similar to those of wild-type HSV-1. We conclude from these studies that phosphorylation is likely a key modulator of ICP0's biological activities in a mouse ocular model of HSV-1 infection.