Intracellular mechanisms of reovirus genome replication and particle assembly

项目概述：本项目的长期、广泛目标是了解细胞内、分子内和细胞外的生物学特性。 哺乳动物正呼肠孤病毒（呼肠孤病毒），这是人类的已知原因， 感染和建立模型的研究病毒的发病机制，介导的复杂，动态 病毒基因组复制、包装和颗粒组装的过程。本提案的具体目标 主要集中在病毒的非结构蛋白muNS，它已被证明在形成中发挥关键作用， 在这些细胞中，呼肠孤病毒复制的许多步骤和 集会发生。相关结构被同一分类系统的其他成员用于类似目的。 多节段双链RNA病毒家族，如人类病原体轮状病毒和 家畜病原体环状病毒拟议的实验将试图表征muNS蛋白的特征， 这决定了它的自我关联能力和与细胞因子的关联，以诱导 在不存在其他呼肠孤病毒蛋白的情况下的工厂样包涵体（Aim 2），以及其缔合能力 与其他病毒蛋白质和潜在的RNA以及招募他们的工厂或工厂样夹杂物 (Aim 1）。其他拟议的实验将试图完善和应用遗传方法，包括RNA 干扰和基于载体的反式互补，用于测试muNS和工厂在 呼肠孤病毒的生产性感染（目的3）。在这些目标的框架内，其他病毒和细胞 建议对与muNS相关的组件进行研究，特别强调 病毒sigmaNS和mu2蛋白，它们也参与了确定 工厂与公共卫生的相关性：病毒基因组复制和颗粒中的细胞内步骤 对于许多病毒来说，组装仍然知之甚少，一旦更好地理解， 许多抗病毒治疗的新靶点。呼肠孤病毒工厂在这方面提供了一个有用的模型。的 本建议关注的一种特殊病毒，哺乳动物呼肠孤病毒，是一个分类学家族的原型 包括人类、食用动物和食用植物的重要病原体，而且正在研究中， 溶瘤剂。

Project Summary: The long-term, broad objective of this project is to understand the intracellular, molecular mechanisms by which mammalian orthoreoviruses (reoviruses), which are known causes of human infections and established models for studies of viral pathogenesis, mediate the complex, dynamic processes of viral genome replication, packaging, and particle assembly. The specific aims of this proposal are focused on the viral non-structural protein muNS, which has been shown to play key roles in forming specialized cytoplasmic structures, the "viral factories", in which many of the steps in reovirus replication and assembly occur. Related structures are used for similar purposes by other members of the same taxonomic family of multi-segmented double-stranded RNA viruses, such as the human pathogen rotavirus and the livestock pathogen orbivirus. Proposed experiments will attempt to characterize features of the muNS protein that determine its capacity for self-association and association with cellular factors to induce the formation of factory-like inclusions in the absence of other reovirus proteins (Aim 2), as well as its capacity for association with other viral proteins and potentially RNAs as well to recruit them to the factories or factory-like inclusions (Aim 1). Other proposed experiments will attempt to refine and apply genetic approaches, including RNA interference and vector-based trans-complementation, for testing the roles of muNS and the factories in productive infections by reoviruses (Aim 3). Within the framework of these aims, other viral and cellular components are proposed to be studied insofar as they associate with muNS, with a special emphasis on the viral sigmaNS and mu2 proteins, which have also been implicated in determining features or functions of the factories. Relevance to public health: The intracellular steps in viral genome replication and particle assembly are still poorly understood for many viruses and are expected, once better understood, to present many new targets for anti-viral therapies. The reovirus factories provide a useful model in this regard. The particular virus on which this proposal focuses, mammalian reovirus, is the prototype of a taxonomic family that includes important pathogens of humans, food animals, and food plants, and is moreover under study as an oncolytic agent.