Alpha-herpevirus assembly egress and viral particle heterogeneity

描述（由申请人提供）：神经侵袭性疱疹病毒是一种高度流行的α -疱疹病毒亚家族，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）。这一组的另一个成员是具有兽医意义的病毒，伪狂犬病毒（PRV），它历来为研究病毒发病机制提供了模型。尽管有抗病毒化合物阿昔洛韦，但由这些病毒引起的几种严重疾病在我国和全世界仍然普遍存在。与高发病率或高死亡率相关的感染包括脑炎、角膜炎、带状疱疹和新生儿的播散性感染。干扰这些病毒的组装和输出的新策略可能对治疗感染有价值，但疱疹病毒感染周期的大部分仍未确定。在这个应用中，我们利用我们在感染性克隆诱变和单病毒颗粒荧光成像方法方面的专业知识来解剖活细胞和细胞外的病毒结构组成，并解决指导病毒组装和出口的分子途径。新的证据表明，非常大的疱疹病毒被膜蛋白VP1/2在感染细胞的细胞核和细胞质中组装的不同阶段是病毒组装的关键效应物。这一建议是基于这样的假设：疱疹病毒的组装和输出是通过感染细胞的细胞核和细胞质中的一系列连续步骤发生的耦合过程，并且这些步骤中的每一个都部分受到VP1/2蛋白的影响。我们的目标是在蛋白质相互作用水平上完善我们对这些步骤的理解，这些步骤有助于病毒输出的动力学。本建议包括对两种神经侵袭性疱疹病毒亚群的模型病毒进行比较研究，即单纯型病毒（以HSV-1为代表）和水痘病毒（以PRV为代表），以对这些病毒的特性进行全面分析。

DESCRIPTION (provided by applicant): 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 worldwide. Infections associated with high rates of morbidity or mortality includes 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 affected 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 simplex viruses (represented by HSV-1) and the varicelloviruses (represented by PRV), to develop a comprehensive analysis of the properties of these viruses.