Patel A H, Rixon F J, Cunningham C, Davison A J
MRC Virololgy Unit, Institute of Virology, Glasgow, United Kingdom.
Virology. 1996 Mar 1;217(1):111-23. doi: 10.1006/viro.1996.0098.
Previous studies have shown that the protein encoded by herpes simplex virus type 1 (HSV-1) gene UL6 is required for processing and packaging of replicated viral DNA and is a minor component of virions and capsids. In this report, we describe the construction of UL6- HSV-1 mutants with a disrupted UL6 gene using complementing cells and show that they fail to synthesize the UL6 protein or produce infectious virus in noncomplementing cells. The mutants synthesized but failed to process and encapsidate viral DNA and accumulated only immature capsids which lacked the UL6 protein. Immunofluorescence analysis showed that the UL6 protein, when expressed transiently in transfected cells in the absence of other HSV-1 proteins, is localized exclusively to the nucleus. We also investigated an HSV-1 mutant with a defect in gene UL33, the product of which is also thought to be involved in viral DNA processing and packaging. The phenotype of this mutant on noncomplementing cells with regard to failure to process and encapsidate viral DNA, accumulation of immature capsids, and inability to produce infectious virus was the same as that of UL6- viruses. This mutant, however, produced capsids containing the UL6 protein, indicating that association of the UL6 protein with the capsid is independent of the UL33 protein.
先前的研究表明，单纯疱疹病毒1型（HSV-1）基因UL6编码的蛋白质是复制的病毒DNA加工和包装所必需的，并且是病毒粒子和衣壳的次要成分。在本报告中，我们描述了使用互补细胞构建具有破坏的UL6基因的UL6-HSV-1突变体，并表明它们在非互补细胞中无法合成UL6蛋白或产生感染性病毒。这些突变体合成了病毒DNA，但无法对其进行加工和包装，仅积累了缺乏UL6蛋白的未成熟衣壳。免疫荧光分析表明，当UL6蛋白在没有其他HSV-1蛋白的情况下在转染细胞中瞬时表达时，它仅定位于细胞核。我们还研究了一个基因UL33有缺陷的HSV-1突变体，其产物也被认为参与病毒DNA的加工和包装。该突变体在非互补细胞上关于无法加工和包装病毒DNA、未成熟衣壳的积累以及无法产生感染性病毒的表型与UL6-病毒相同。然而，该突变体产生了含有UL6蛋白的衣壳，表明UL6蛋白与衣壳的结合独立于UL33蛋白。
