Alpha-herpevirus assembly egress and viral particle heterogeneity

描述(申请人提供)：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具有兽医意义的伪狂犬病病毒(PRV)，它历史上曾为研究病毒的发病机制提供模型。尽管有抗病毒化合物阿昔洛韦可用，但由这些病毒引起的几种严重形式的疾病仍然在该国和世界各地流行。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输出的新策略可能被证明对感染的治疗有价值，但疱疹病毒感染周期的大部分仍未确定。在这项应用中，我们利用我们在感染性克隆突变和单病毒颗粒荧光成像方法方面的专业知识来剖析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器。这一建议是基于这样的假设，即疱疹病毒的组装和出口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步都部分受到VP1/2蛋白的影响。我们的目标是在蛋白质相互作用的水平上完善我们对这些步骤的理解，这些相互作用有助于病毒出口的动态。这项建议包括对神经侵袭性疱疹病毒的两个亚组--单纯疱疹病毒(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，以开发对这些病毒特性的全面分析。 公共卫生相关性：神经侵袭性疱疹病毒是许多严重疾病的病原体，包括带状疱疹、脑炎、新生儿感染和疱疹角膜炎(在美国和其他工业化国家，疱疹是感染性失明的主要原因)。这项建议侧重于了解疱疹病毒颗粒组装和排出的分子机制，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.