Phosphorylation of structural components promotes dissociation of the herpes simplex virus type 1 tegument
Phosphorylation of structural components promotes dissociation of the herpes simplex virus type 1 tegument
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DOI：
10.1128/jvi.72.9.7108-7114.1998
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发表时间：
1998-09-01
影响因子：
5.4
通讯作者：
Meredith, DM
中科院分区：
文献类型：
--
作者：
Morrison, EE;Wang, YF;Meredith, DM
利用体外试验研究了磷酸化在单纯疱疹病毒1型（HSV-1）被膜结构组分解离中的作用。在去污剂存在下，向野生型病毒体中加入生理浓度的ATP和镁促进了VP 13/14和VP 22的释放。VP 1/2和UL 13蛋白激酶没有显著溶解。然而，使用具有灭活的UL 13蛋白的病毒，我们发现VP 22的释放严重受损。将酪蛋白激酶II（CKII）添加到UL 13突变体病毒粒子中促进VP 22的释放。热灭活的病毒粒子或磷酸酶抑制这两种蛋白质的释放。将放射性标记的ATP掺入试验中证明了VP 1/2、VP 13/14、VP 16和VP 22的磷酸化。用重组激酶孵育经去污剂纯化的热灭活的衣壳-被膜，搁置了由CKII引起的VP 1/2磷酸化，由CKII、蛋白激酶A（PKA）和PKC引起的VP 13/14磷酸化，由PKA引起的VP 16磷酸化，以及由CKII和PKC引起的VP 22磷酸化。磷酸化VP 22的蛋白水解图谱和磷酸化氨基酸分析与先前发表的工作相关。在放线菌酮存在下感染的细胞中证实了病毒体相关的VP 13/14、VP 16和VP 22的磷酸化。在体外释放试验中使用马疱疹病毒1导致VP 10（HSV-1 VP 13/14的同源物）的释放增强。这些结果表明，在感染的细胞中，从α疱疹病毒病毒粒子的主要被膜蛋白的解离可能是由病毒粒子相关和细胞激酶介导的磷酸化事件启动。
The role of phosphorylation in the dissociation of structural components of the herpes simplex virus type 1 (HSV-1) tegument was investigated, using an in vitro assay. Addition of physiological concentrations of ATP and magnesium to wild-type virions in the presence of detergent promoted the release of VP13/14 and VP22. VP1/2 and the UL13 protein kinase were not significantly solubilized. However, using a virus with an inactivated UL13 protein, we found that the release of VP22 was severely impaired. Addition of casein kinase II (CKII) to UL13 mutant virions promoted VP22 release. Heat inactivation of virions or addition of phosphatase inhibited the release of both proteins. Incorporation of radiolabeled ATP into the assay demonstrated the phosphorylation of VP1/2, VP13/14, VP16, and VP22. Incubation of detergent-purified, heat-inactivated capsid-tegument with recombinant kinases shelved VP1/2 phosphorylation by CKII, VP13/14 phosphorylation by CKII, protein kinase A (PKA), and PKC, VP16 phosphorylation by PKA, and VP22 phosphorylation by CKII and PKC. Proteolytic mapping and phosphoamino acid analysis of phosphorylated VP22 correlated with previously published work The phosphorylation of virion-associated VP13/14, VP16, and VP22 was demonstrated in cells infected in the presence of cycloheximide. Use of equine herpesvirus 1 in the in vitro release assay resulted in the enhanced release of VP10, the homolog of HSV-1 VP13/14. These results suggest that the dissociation of major tegument proteins from alphaherpesvirus virions in infected cells may be initiated by phosphorylation events mediated by both virion-associated and cellular kinases.