Alpha-herpesvirus assembly, egress and viral particle heterogeneity

神经侵袭性疱疹病毒是α-疱疹病毒亚科的高度流行的组， 包括人类病原体：单纯疱疹病毒1型和2型（HSV-1，HSV-2），以及水痘 带状疱疹病毒（VZV）。该组的另一个成员是具有兽医意义的病毒， 伪狂犬病病毒（PRV），其在历史上为研究病毒致病机制提供了模型。 尽管抗病毒化合物阿昔洛韦的可用性，几种严重形式的疾病引起的 这些病毒在这个国家和世界范围内仍然流行。与高发病率相关的感染 发病率或死亡率包括脑炎、角膜炎、带状疱疹和新生儿的播散性感染。 干扰这些病毒组装和外出的新策略可能被证明是有价值的， 虽然疱疹病毒感染的治疗已经取得了进展，但疱疹病毒感染的大部分周期仍不明确。在这 我们利用我们在感染性克隆诱变和单病毒颗粒方面的专业知识， 荧光成像方法解剖活细胞和细胞外的病毒结构组成， 并解决引导病毒组装和外出的分子途径。新的证据提供了 这表明疱疹病毒被膜蛋白VP 1/2是病毒装配的关键效应子 在受感染细胞的细胞核和细胞质中的不同组装阶段。这项建议是 基于疱疹病毒装配和外出是发生在 在感染细胞的细胞核和细胞质中通过一系列顺序步骤， 这些步骤的一部分是由VP 1/2蛋白的影响。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，有助于病毒外出的动力学。这 该提案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 亚组，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的分析。

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.