Virus-host interactions in replication complex assembly

病毒的致病机制与动态和复杂的宿主-病原体相互作用密切相关。这个 这些基本相互作用的机制，特别是具有相似基因组结构的病毒共享的机制，代表了抗病毒药物的诱人潜在靶点。包含阳性单链RNA基因组的病毒，如丙型肝炎病毒、西尼罗河病毒和SARS冠状病毒，代表了导致重大人类疾病的各种病原体，但对这些疾病几乎没有有效的治疗方法。尽管这些病毒引起了不同的临床症状，但所有具有特征的正链RNA病毒都利用细胞内膜形成和功能病毒RNA复制复合体。然而，病毒RNA复制复合体在特定的细胞内组装的机制 人们对细胞膜的了解还不够深入。该项目的长期目标是阐明促进正链RNA病毒复制复合体组装和功能的宿主-病原体相互作用。对这些相互作用的详细了解将有助于深入了解病毒的发病机制，并有可能确定广泛有效的抗病毒药物的新靶点。这项提案的具体重点是定义病毒RNA复制复合体组装中的早期事件。病毒编码的RNA复制复合体蛋白的靶向、转运和与细胞内膜的初始相互作用是复制复合体组装的重要步骤，因此是病毒致病的重要决定因素。这个 这项研究的总体策略是利用鸡舍病毒这一已建立的、通用的模型来研究正链RNA病毒的结构和复制，以探讨病毒RNA复制复合体组装的机制。生物化学、分子和遗传学方法将被用来研究鸡舍病毒依赖于RNA的RNA聚合酶的靶向和细胞内膜的运输。这项建议的具体目的是为了实现两个目标：1)了解细胞伴侣蛋白在鸡舍病毒RNA复制复合体组装中的作用；2)确定负责鸡舍病毒RNA复制复合体细胞内定位的膜受体。

Viral pathogenesis is intimately linked with dynamic and complex host-pathogen interactions. The mechanisms underlying these essential interactions, in particular those shared by viruses with similar genomic structures, represent attractive potential targets for antiviral drugs. Viruses that contain a positive-sense single-stranded RNA genome, such as hepatitis C virus, West Nile virus, and the SARS coronavirus, represent a diverse group of pathogens responsible for significant human diseases for which few effective therapies exist. Despite the varied clinical syndromes caused by these viruses, all characterized positive-strand RNA viruses use intracellular membranes for viral RNA replication complex formation and function. However, the mechanisms whereby viral RNA replication complexes assemble on specific intracellular membranes are not well understood. The long-term objectives of this project are to elucidate the host-pathogen interactions that facilitate positive-strand RNA virus replication complex assembly and function. A detailed understanding of these interactions will provide insight into the mechanisms of viral pathogenesis, and potentially identify novel targets for broadly effective antiviral drugs. The specific focus of this proposal is to define the early events in viral RNA replication complex assembly. The targeting, transport, and initial interactions of virus-encoded RNA replication complex proteins with intracellular membranes are essential steps in replication complex assembly, and therefore are important determinants of viral pathogenesis. The general strategy of the proposed research is to use Flock house virus, an established and versatile model used to study positive-strand RNA virus structure and replication, to investigate the mechanisms of viral RNA replication complex assembly. Biochemical, molecular, and genetic approaches will be used to investigate the targeting and transport of the Flock house virus RNA-dependent RNA polymerase to intracellular membranes. The specific aims of this proposal are designed to accomplish two goals: 1) understand the role of cellular chaperone proteins in Flock house virus RNA replication complex assembly; and 2) define the membrane receptor responsible for Flock house virus RNA replication complex intracellular localization.