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α-疱疹病毒亚科的一个高度流行的组，包括人类病原体：单纯疱疹病毒1型和2型（HSV-1、HSV-2）和水痘带状疱疹病毒（VZV）。该组的另一个成员是具有兽医学意义的病毒，伪狂犬病病毒（PRV），其在历史上提供了研究病毒发病机制的模型。尽管抗病毒化合物阿昔洛韦的可用性，由这些病毒引起的几种严重形式的疾病仍然在这个国家和世界范围内流行。与高发病率或死亡率相关的感染包括脑炎、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外出的新策略可能对治疗感染有价值，但疱疹病毒的感染周期仍不明确。在此应用中，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，剖析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。提供了新的证据表明，非常大的疱疹病毒被膜蛋白，VP 1/2，是一个关键的效应，在不同阶段的组装在感染细胞的细胞核和细胞质中的病毒组装。该建议是基于这样的假设，即疱疹病毒组装和出口是通过一系列连续步骤在感染细胞的细胞核和细胞质中发生的耦合过程，并且这些步骤中的每一个都部分地受到VP 1/2蛋白的影响。我们的目标是在蛋白质相互作用的水平上完善我们对这些步骤的理解，这些步骤有助于病毒流出的动力学。该提案包括对来自两个神经侵袭性疱疹病毒亚组（单纯病毒（以HSV-1为代表）和水痘病毒（以PRV为代表））的模型病毒进行比较研究，以全面分析这些病毒的特性。 公共卫生相关性：神经侵入性疱疹病毒是许多严重疾病的病原体，包括带状疱疹、脑炎、新生儿感染和疱疹性角膜炎（在美国和其他工业化国家中感染性失明的主要原因）。该提案的重点是了解疱疹病毒颗粒组装和外出的分子机制，长期目标是确定干预疾病进展的新靶点。

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. PUBLIC HEALTH RELEVANCE: Neuroinvasive herpesviruses are the causative agents of a number of severe diseases including shingles, encephalitis, neonatal infections and herpes keratitis (the leading cause of infectious blindness in the USA and other industrialized nations). This proposal focuses on understanding the molecular mechanisms that underlie the assembly and egress of herpesvirus particles, with the long- term goal of identifying new targets for the intervention of disease progression.