UL33 GENE PRODUCT OF HSV1

在单纯疱疹病毒（HSV-1）感染细胞的细胞核内，复制的HSV-DNA形成首尾相连的连接体，这些连接体被切割成基因组长度的分子，随后易位到执行衣壳中。与此建议相关的长期目标是定义HSV DNA切割和包装的分子机制。HSV基因UL6、UL15、UL17、UL28、UL32和UL33在衣壳组装中是必不可少的，但已被证明是病毒DNA切割和包装所必需的。UL33被输入到感染细胞的细胞核中。输入是由晚期基因产物介导的。本提案的总体目标是通过1)鉴定病毒UL33核重要基因产物和2)鉴定UL33与其他HSV蛋白之间蛋白-蛋白相互作用所需的关键蛋白结构域来表征UL33蛋白在HSV-1包装体中的作用。我们对UL33基因产物进行了外视标记，并开发了一种核输入试验，检测缺乏特定切割/包装基因的病毒对UL33蛋白核输入的影响。

DESCRIPTION Within the nuclei of herpes simplex virus (HSV-1) infected cells, replicating HSV-DNA forms head-to-tail concatemers which are cleaved to genome length molecules and subsequently translocated into performed capsids. The long term objective related to this proposal is the definition of the molecular mechanisms of HSV DNA cleavage and packaging. The HSV genes UL6, UL15, UL17, UL28, UL32, and UL33 are dispensable for capsid assembly, but have been shown to be necessary for viral DNA cleavage and packaging. UL33 is imported into the nucleus in infected cells. Importation is mediated by late gene product(s). The overall aim of this proposal is to characterize the role of UL33 protein in the HSV-1 packasome by 1) Identifying gene products required for viral UL33 nuclear important and 2) Identifying the critical protein domains necessary for protein-protein interactions between UL33 and other HSV proteins. We have eptiopically-tagged the UL33 gene product and developed a nuclear importation assay examining the effect of viruses lacking specific cleavage/packaging genes on nuclear importation of UL33 protein.