HSV 1 PROCESSING/PACKAGING GENES

单纯疱疹病毒在人群中是地方性的，并导致各种临床疾病，其中一些危及生命，特别是在免疫功能低下的个人或新生儿中。对病毒突变体的分析表明，病毒基因组的成熟和囊化过程似乎是一个多步骤的操作，涉及至少8种病毒基因产物的功能。这些病毒基因(UL6、UL12、UL15、ULI7、UL25、UL28、UL32和UL33)在疱疹病毒家族中保存良好，表明它们可能在病毒生命周期中发挥重要作用。这项建议的目的是通过遗传和生化分析更好地了解基因组加工和打包的事件。第一个目的是确定UL15和UL28在卵裂和包装中的作用。我们计划测试这些蛋白质编码假定的终止酶/转位酶的假设。为此，我们将尝试纯化可溶的UL15/UL28蛋白复合体，并对终止酶的预期功能进行生化测试。我们还将扩大对这两种基因产品的基因分析。第二个目标是进一步确定UL6和UL32的作用。初步证据表明，UL6可能是一种门户蛋白，UL32是衣壳蛋白在感染细胞中正确定位所必需的。这些任务将通过基因分析进行测试。第三个目标是继续鉴定与Proapsids相关的切割和包装蛋白，这些蛋白是从感染了在包埋过程中不同阶段被阻断的突变体的细胞中分离出来的。预计这种类型的分析将提供关于这些蛋白质作用的更多线索。我们还计划检查衣壳在感染过程中的定位，以表征包膜过程的空间组织。长期目标包括发展体外分裂和包装反应。预计这一建议不仅将加深我们对基因组成熟和包囊化机制的理解，还可能导致抗病毒治疗新策略的发展。

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.