Functions of the ORF9-to-ORF12 gene cluster in varicella-zoster virus replication and in the pathogenesis of skin infection
Functions of the ORF9-to-ORF12 gene cluster in varicella-zoster virus replication and in the pathogenesis of skin infection
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DOI：
10.1128/jvi.00303-08
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发表时间：
2008-06-01
影响因子：
5.4
通讯作者：
Arvin, Ann M.
中科院分区：
文献类型：
--
作者：
Che, Xibing;Reichelt, Mike;Arvin, Ann M.
由水痘带状疱疹病毒 (VZV) 开放阅读框 9 (OPF9) 至 ORF12 组成的基因簇编码四种假定的被膜蛋白，并且在大多数α疱疹病毒中高度保守。在这些实验中，该簇内的基因从 VZV 亲本 Oka (POKA) 中单独或组合删除，并用体外培养细胞和 SCID 小鼠体内的人皮肤异种移植物评估 VZV 复制的后果。正如 ORF10 的报道，ORF11 和 ORF12 对于 VZV 在黑色素瘤和人胚胎成纤维细胞中的复制来说是可有可无的。相反，ORF9的删除与感染性病毒的恢复不相容。 ORF9 定位于病毒颗粒外皮，并与 VZV 感染细胞中的必需蛋白——糖蛋白 E 形成复合物。缺乏 ORF10 和 ORF11 (POKA Delta 10/11)、ORF11 和 ORF12 (POKA Delta 11/12) 或 ORF10、ORF11 和 ORF12 (POKA Delta 10/11/12) 的重组体在培养细胞中可存活。它们的生长动力学与 POKA 没有什么不同，核衣壳的形成和病毒颗粒的组装也没有受到干扰。此外，这些缺失突变体与 POKA 相比，在原代人扁桃体 T 细胞的感染水平上没有表现出差异。 ORF12的删除对皮肤感染没有影响，而POKA Delta 11、POKA Delta 10/11和POKA Delta 11/12的复制严重减少，并且从接种POKA Delta 10/11/12的皮肤异种移植物中没有回收到病毒。这些结果表明，除ORF9外，ORF9至ORF12基因簇内的各个基因是可有可无的，可以同时删除，而不会对VZV体外复制产生任何明显影响，但ORF10至ORF12基因簇对于VZV体内皮肤毒力至关重要。
The gene cluster composed of varicella-zoster virus (VZV) open reading frame 9 (OPF9) to ORF12 encodes four putative tegument proteins and is highly conserved in most alphaherpesviruses. In these experiments, the genes within this cluster were deleted from the VZV parent Oka (POKA) individually or in combination, and the consequences for VZV replication were evaluated with cultured cells in vitro and with human skin xenografts in SCID mice in vivo. As has been reported for ORF10, ORF11 and ORF12 were dispensable for VZV replication in melanoma and human embryonic fibroblast cells. In contrast, deletion of ORF9 was incompatible with the recovery of infectious virus. ORF9 localized to the virion tegument and formed complexes with glycoprotein E, which is an essential protein, in VZV-infected cells. Recombinants lacking ORF10 and ORF11 (POKA Delta 10/11), ORF11 and ORF12 (POKA Delta 11/12), or ORF10, ORF11 and ORF12 (POKA Delta 10/11/12) were viable in cultured cells. Their growth kinetics did not differ from those of POKA, and nucleocapsid formation and virion assembly were not disrupted. In addition, these deletion mutants showed no differences compared to POKA in infectivity levels for primary human tonsil T cells. Deletion of ORF12 had no effect on skin infection, whereas replication of POKA Delta 11, POKA Delta 10/11, and POKA Delta 11/12 was severely reduced, and no virus was recovered from skin xenografts inoculated with POKA Delta 10/11/12. These results indicate that with the exception of ORF9, the individual genes within the ORF9-to-ORF12 gene cluster are dispensable and can be deleted simultaneously without any apparent effect on VZV replication in vitro but that the ORF10-to-ORF12 cluster is essential for VZV virulence in skin in vivo.