Herpes Simplex Virus Capsid Assembly and DNA Packaging

人类疱疹病毒感染在人群中是地方性的，一些疱疹病毒引起严重的 疾病，特别是免疫受损的个体和新生儿。免疫功能低下的增加 人口的增加产生了对针对疱疹病毒的抗病毒药物的未满足的医疗需求。建议的目标 一个项目是为了更好地了解单纯疱疹病毒（HSV）中的DNA切割和包装 通过分析这一过程所需的两种病毒蛋白质UL 25和UL 28。具体 这项研究背后的假设是，UL 25和UL 28是将DNA包装到HSV中所必需的 在裂解包装反应中的不同阶段分离衣壳。UL 28蛋白是切割 UL 25可以将多联体DNA转化为单位长度的分子，而UL 25在产生多联体DNA中起重要作用。 含有DNA的衣壳。虽然重点将放在UL 25和UL 28上，但目标是表征 切割/包装过程，使用遗传和生物化学方法来检查突变是如何在 UL 25和UL 28基因影响这些蛋白质与以下物质的相互作用：（i）DNA，（ii）衣壳蛋白，（iii） 切割/包装蛋白和（iv）宿主细胞蛋白。该项目的具体目标是：1。确定 UL 25的衣壳位置、DNA结合特异性和蛋白质-蛋白质相互作用。2.识别功能 UL 28的结构域对于衣壳掺入、结合病毒DNA包装位点和 通过表征一系列HSV-1 UL 28突变体与末端酶（UL 15）和门蛋白（UL 6）的相互作用 我们已经分离出来了;并绘制第二个位点突变的致命UL 28接头之一的回复体- 插入突变体，因为标记拯救和DNA测序已经证明突变不 与UL 28基因相关。3.检测UL 25和UL 28与病毒和细胞蛋白的相互作用， 蛋白质组学和生物化学方法，以确定必要的蛋白质-蛋白质相互作用和蛋白质 参与切割/包装反应的复合物。这些研究旨在阐明 HSVDNA切割和包装的潜在机制，并可能为HSVDNA切割和包装提供新的靶点。 开发抗病毒药物。

Infections with human herpesviruses are endemic in the population with some herpesviruses causing severe disease, especially in immune impaired individuals and newborns. The increased immunocompromised population has created an unmet medical need for antivirals against herpesviruses. The goal of the proposed project is to gain a better understanding of DNA cleavage and packaging in herpes simplex virus (HSV) through the analysis of two viral proteins, UL25 and UL28, that are required for this process. The specific hypothesis behind the proposed research is that UL25 and UL28 are required for packaging DNA into HSV capsids at separate stages in the cleavage packaging reaction. The UL28 protein is required for cleavage of concatemeric DNA into unit length molecules while UL25 serves an essential function in the production of DNA containing capsids. Although, the focus will be on UL25 and UL28 the goal is to characterize the cleavage/packaging process using genetic and biochemical approaches to examine how mutations in the UL25 and UL28 genes affect the interaction of these proteins with: (i) DNA, (ii) capsid proteins, (iii) cleavage/packaging proteins and (iv)host cell proteins. Specific goals of the project are to: 1. Determine the capsid location, DNA binding specificity and protein-protein interactions of UL25. 2. Identify functional domains of UL28 that are important for capsid incorporation, binding viral DNA packaging sites and for interaction with terminase (UL15) and portal protein (UL6) by characterizing a series of HSV-1 UL28 mutants that we have isolated; and map the second site mutation for a revertent of one of the lethal UL28 linker- insertion mutants since marker rescue and DNA sequencing have demonstrated that the mutation does not map to the UL28 gene. 3. Examine the interaction of UL25 and UL28 with viral and cell proteins using proteomic and biochemical approaches in order to identify essential protein-protein interactions and protein complexes that are involved in the cleavage/packaging reaction. These studies aim to elucidate the mechanism underlying HSV DNA cleavage and packaging and may suggest novel targets for the development of antivirals.