Alpha-herpesvirus assembly, egress and viral particle heterogeneity

神经侵袭性疱疹病毒是阿尔法疱疹病毒亚家族中高度流行的一组病毒， 包括人类病原体：单纯疱疹病毒1型和2型(HSV-1、HSV-2)和水痘 带状疱疹病毒(VZV)。这个小组的另一个成员是一种具有兽医意义的病毒， 伪狂犬病病毒(PRV)，历史上为研究病毒的致病机理提供了模型。 尽管有抗病毒化合物阿昔洛韦的供应，但由以下原因引起的几种严重疾病 这些病毒仍然在这个国家和世界范围内流行。与高发病率相关的感染 发病率或死亡率包括脑炎、角膜炎、带状疱疹和新生儿播散性感染。 干扰这些病毒的组装和出口的新策略可能被证明是有价值的 关于感染的治疗，然而疱疹病毒感染周期的大部分仍未确定。在这 应用我们利用我们在感染性克隆诱变和单个病毒颗粒方面的专业知识 荧光成像方法剖析活细胞内和细胞外的病毒结构成分， 并解决引导病毒组装和出口的分子途径。提供了新的证据 表明疱疹病毒被膜蛋白VP1/2是病毒组装的关键效应器 在感染细胞的细胞核和细胞质中组装的不同阶段。这项建议是 基于疱疹病毒组装和出口是发生的耦合过程的假设 通过感染细胞的细胞核和细胞质中的一系列顺序步骤，每个步骤 这些步骤中的一部分是受VP1/2蛋白影响的。我们的目标是完善我们对这些问题的理解 在蛋白质相互作用的水平上的步骤，有助于病毒出口的动态。这 建议包括对两种神经侵袭性疱疹病毒的模型病毒进行比较研究 亚群，单纯疱疹病毒(以HSV-1为代表)和水痘病毒(以PRV为代表)， 对这些病毒的特性进行全面分析。

The neuroinvasive herpesviruses are a highly-prevalent group of the alpha-herpesvirus subfamily that includes the human pathogens: herpes simplex virus types 1 and 2 (HSV-1, HSV-2), and varicella zoster virus (VZV). An additional member of this group is a virus of veterinary significance, pseudorabies virus (PRV), which historically has provided models for studying viral pathogenesis. Despite the availability of the antiviral compound acyclovir, several severe forms of disease caused by these viruses remain prevalent in this country and world wide. Infections associated with high rates of morbidity or mortality include encephalitis, keratitis, shingles and disseminated infections in newborns. Novel strategies to interfere with the assembly and egress of these viruses could prove valuable to treatment of infections, yet much of the herpesvirus infectious cycle remains undefined. In this application we leverage our expertise in infectious clone mutagenesis and single viral particle fluorescence imaging methods to dissect viral structural composition in living-cells and extracellularly, and to address the molecular pathways guiding viral assembly and egress. New evidence is provided indicating that the very large herpesvirus tegument protein, VP1/2, is a key effector of viral assembly during distinct stages of assembly in the nucleus and cytoplasm of infected cells. This proposal is based on the hypothesis that herpesvirus assembly and egress are coupled processes that occur through a series of sequential steps both in the nucleus and cytoplasm of infected cells, and that each of these steps are effected in part by the VP1/2 protein. Our goal is to refine our understanding of these steps at the level of the protein interactions that contribute to the dynamics of viral egress. This proposal includes comparative studies of model viruses from the two neuroinvasive herpesviruses subgroups, the simplexviruses (represented by HSV-1) and the varicelloviruses (represented by PRV), to develop a comprehensive analysis of the properties of these viruses.