The Cell Biology of Coronavirus Infection

冠状病毒是引起人类呼吸道感染的正链RNA病毒， 其他动物重要胃肠和神经系统疾病。小鼠肝炎病毒（MHV）是一种 用于研究冠状病毒复制和发病机制的模型。拟议研究的中心目标是 定义小鼠肝炎病毒在病毒复制形成和功能过程中与宿主细胞的相互作用 配合物MHV指导病毒RNA在膜复制复合物上合成， throughout the cytoplasmic细胞质.这些复合物与中间体中病毒粒子组装的位点不同 高尔基体（ERGIC）。由复制酶基因多蛋白加工而成的蛋白质定位于 核衣壳（N）蛋白在两个密切相关的蛋白质复合物中的病毒RNA合成位点， p22/p65复合物和hel/N复合物。在感染的晚期，hel/N复合物从细胞中分离。 p22/p65复合物并易位到ERGIC中的病毒体组装位点。然而，精确 复制复合物的组成尚不清楚，并且hel/N复合物易位的机制已被证实。 尚未确定。该提案描述了一个综合研究计划，以确定组织和 MHV复制复合物的易位。在具体目标1中， 将测定MHV复制复合物。在具体目标2中，研究了hel/N络合物的作用机理， 将定义向ERGIC的易位。在特定目标3中，病毒蛋白在复制中的功能 将确定复杂易位。这些研究将确定基本机制， MHV指导复制复合物的形成和功能，并将定义病毒-细胞相互作用， 是病毒成功复制的关键。

Coronaviruses are positive-strand RNA viruses that cause respiratory infections in humans and important gastrointestinal and neurologic disease in other animal species. Mouse hepatitis virus (MHV) is a model for studies of coronavirus replication and pathogenesis. The central goal of the proposed research is to define interactions of mouse hepatitis virus with host cells during formation and function of viral replication complexes. MHV directs viral RNA synthesis on membranous replication complexes that are distributed thoughout the cytoplasm. These complexes are distinct from sites ofvirion assembly in the intermediate compartment of the Golgi (ERGIC). Proteins processed from the replicase gene polyprotein localize with the nucleocapsid (N) protein at sites of viral RNA synthesis in two closely associated protein complexes, the p22/p65 complex and the hel/N complex. At late times of infection, the hel/N complexes separate from the p22/p65 complexes and translocate to sites of virion assembly in the ERGIC. However, the precise composition of replication complexes is not known, and the mechanism of hel/N complex translocation has not been determined. This proposal describes an integrated research program to define the organization and translocation of MHV replication complexes. In Specific Aim 1, the protein and membrane composition of MHV replication complexes will be determined. In Speeific Aim 2, the mechanism of hel/N complex translocation to the ERGIC will be defined. In Specific Aim 3, the functions of viral proteins in replication complex translocation will be determined. These studies will identify fundamental mechanisms by which MHV directs replication complex formation and function and will define virus-cell interactions that are essential for successful virus replication.