Roles of Us8A and Its Phosphorylation Mediated by Us3 in Herpes Simplex Virus 1 Pathogenesis
Roles of Us8A and Its Phosphorylation Mediated by Us3 in Herpes Simplex Virus 1 Pathogenesis
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DOI：
10.1128/jvi.00446-16
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发表时间：
2016-06-01
影响因子：
5.4
通讯作者：
Kawaguchi, Yasushi
中科院分区：
文献类型：
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作者：
Kato, Akihisa;Ando, Tomoko;Kawaguchi, Yasushi
单纯疱疹病毒1（HSV-1）Us 8A基因与编码糖蛋白E（gE）的基因重叠。以前的研究已经使用Us 8A和gE中的无效突变研究了Us 8A在HSV-1感染中的作用;因此，Us 8A的作用仍有待阐明。在这项研究中，我们研究了Us 8A的功能及其在丝氨酸61（Ser-61），我们最近确定为一个磷酸化位点的HSV-1感染的细胞，在HSV-1的发病机制，基于质谱的磷酸化蛋白质组学分析磷酸化。我们观察到（i）感染细胞中Us 8A Ser-61的磷酸化依赖于病毒编码的Us 3蛋白激酶的活性;（ii）与修复的病毒相比，Us 8A无效突变病毒在颅内感染后小鼠中枢神经系统（CNS）中表现出10倍的50%致死量增加;（iii）用丙氨酸（S61 A）替换Us 8A中的Ser-61对颅内感染后小鼠CNS中的毒力几乎没有影响，而其显著降低眼部感染后小鼠的死亡率至与Us 8A无效突变病毒相似的水平;（iv）Us 8A S61 A突变还显著降低眼部感染后小鼠中的病毒产量，主要是三叉神经节和脑中的病毒产量;和（v）Us 8A Ser-61处的拟磷酸化突变恢复了野生型病毒产量和毒力。总的来说，这些结果表明，Us 8A是一种新的HSV-1毒力因子，并建议Us 3介导的磷酸化Us 8A Ser-61调节Us 8A功能的病毒入侵到中枢神经系统从外周sites.IMPORTANCE的DNA基因组内的亚科α疱疹病毒分为独特的长（UL）和独特的短（US）区域。美国地区含有α疱疹病毒特异性基因。最近，对HSV-1眼部分离株的高通量测序显示，Us 8A是定位于Us区域的13个单纯疱疹病毒1（HSV-1）基因中最高度保守的，这表明Us 8A可能在HSV-1生命周期中起重要作用。然而，Us 8A在HSV-1感染中的具体作用仍有待阐明。在这里，我们表明，Us 8A是一个毒力因子HSV-1感染小鼠，和Us 8A的功能，从外周网站的病毒入侵到中枢神经系统的调节由Us 3介导的蛋白质在Ser-61磷酸化。这是首次报道Us 8A及其调控在HSV-1感染中的意义的研究。
The herpes simplex virus 1 (HSV-1) Us8A gene overlaps the gene that encodes glycoprotein E (gE). Previous studies have investigated the roles of Us8A in HSV-1 infection using null mutations in Us8A and gE; therefore, the role of Us8A remains to be elucidated. In this study, we investigated the function of Us8A and its phosphorylation at serine 61 (Ser-61), which we recently identified as a phosphorylation site by mass spectrometry-based phosphoproteomic analysis of HSV-1-infected cells, in HSV-1 pathogenesis. We observed that (i) the phosphorylation of Us8A Ser-61 in infected cells was dependent on the activity of the virus-encoded Us3 protein kinase; (ii) the Us8A null mutant virus exhibited a 10-fold increase in the 50% lethal dose for virulence in the central nervous system (CNS) of mice following intracranial infection compared with a repaired virus; (iii) replacement of Ser-61 with alanine (S61A) in Us8A had little effect on virulence in the CNS of mice following intracranial infection, whereas it significantly reduced the mortality of mice following ocular infection to levels similar to the Us8A null mutant virus; (iv) the Us8A S61A mutation also significantly reduced viral yields in mice following ocular infection, mainly in the trigeminal ganglia and brains; and (v) a phosphomimetic mutation at Us8A Ser-61 restored wild-type viral yields and virulence. Collectively, these results indicate that Us8A is a novel HSV-1 virulence factor and suggest that the Us3-mediated phosphorylation of Us8A Ser-61 regulates Us8A function for viral invasion into the CNS from peripheral sites.IMPORTANCE The DNA genomes of viruses within the subfamily Alphaherpesvirinae are divided into unique long (UL) and unique short (Us) regions. Us regions contain alphaherpesvirus-specific genes. Recently, high-throughput sequencing of ocular isolates of HSV-1 showed that Us8A was the most highly conserved of 13 herpes simplex virus 1 (HSV-1) genes mapped to the Us region, suggesting Us8A may have an important role in the HSV-1 life cycle. However, the specific role of Us8A in HSV-1 infection remains to be elucidated. Here, we show that Us8A is a virulence factor for HSV-1 infection in mice, and the function of Us8A for viral invasion into the central nervous system from peripheral sites is regulated by Us3-mediated phosphorylation of the protein at Ser-61. This is the first study to report the significance of Us8A and its regulation in HSV-1 infection.