HSV 1 PROCESSING/PACKAGING GENES

单纯疱疹病毒在人群中是地方性的，并且引起多种临床疾病，其中一些是危及生命的，特别是在免疫功能低下的个体或新生儿中。 病毒基因组成熟和突变的过程似乎是一个多步骤的操作，涉及至少8个病毒基因产物的功能，如病毒突变体的分析所揭示的。这些病毒基因（UL 6、UL 12、UL 15、UL 17、UL 25、UL 28、UL 32和UL 33）在疱疹病毒家族中非常保守，表明它们可能在病毒生命周期中发挥重要作用。该提案的目的是通过遗传和生化分析更好地了解基因组加工和包装事件。第一个目的是确定UL 15和UL 28在切割和包装中的作用。我们计划测试的假设，这些蛋白质编码假定的终止酶/转位酶。为此，我们将尝试纯化可溶性UL 15/UL 28蛋白复合物，并进行生物化学测试，以确定末端酶的预期功能。我们还将扩展我们对这两种基因产物的遗传分析。第二个目的是进一步表征UL 6和UL 32的作用。初步证据表明，UL 6可能作为一个门户蛋白和UL 32所需的衣壳正确定位在感染的细胞。这些任务将通过遗传分析进行测试。第三个目标是继续鉴定与原衣壳相关的切割和包装蛋白，所述原衣壳分离自在胞苷化过程的各个阶段被阻断的突变体感染的细胞。预计这种类型的分析将提供有关这些蛋白质作用的额外线索。我们还计划检查感染过程中的衣壳定位，以表征衣壳化过程的空间组织。长期目标包括开发体外裂解和包装反应。预计这一提议不仅将增强我们对基因组成熟和失活机制的理解，而且还可能导致开发新的抗病毒治疗策略。

Herpes simplex viruses are endemic in the population and are responsible for a variety of clinical diseases some of which are life threatening, especially in immunocompromised individuals or in newborns. The processes of viral genome maturation and encapsidation appear to be a multistep operation involving the function of at least eight viral gene products as revealed by the analysis of viral mutants. These viral genes (UL6, UL12, UL15, ULI7, UL25, UL28, UL32 and UL33) are well conserved within the herpesvirus family indicating that they likely play important roles in the viral life cycle. The objective of this proposal is to provide a better understanding of the events of genome processing and packaging through genetic and biochemical analysis. The first aim is to determine the role of UL15 and UL28 in cleavage and packaging. We plan to test the hypothesis that these proteins encode the putative terminase/translocase. To this end we will attempt to purify a soluble UL15/UL28 protein complex and test biochemically for functions expected of a terminase. We also will extend our genetic analysis of these two gene products. The second aim is to further characterize the roles of UL6 and UL32. Preliminary evidence suggests that UL6 may act as a portal protein and the UL32 is required for capsids to localize correctly in infected cells. These assignments will be tested by genetic analysis. The third aim is to continue the identification of the cleavage and packaging proteins that are associated with procapsids isolated from cells infected with mutants blocked at various stages in the encapsidation process. It is anticipated that this type of analysis will provide additional clues about the roles of these proteins. We also plan to examine capsid localization during infection to characterize the spatial organization of the encapsidation process. Long term aims include the development of an in vitro cleavage and packaging reaction. It is anticipated that this proposal will not only enhance our understanding of the mechanisms of genome maturation and encapsidation but may also lead to the development of novel strategies for antiviral therapy.