Intracellular mechanisms of reovirus genome replication and particle assembly

呼肠孤病毒基因组复制和颗粒组装的细胞内机制

• 批准号: 7486523
• 负责人: MAX L. NIBERT
• 金额: $ 33.8万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-15 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7486523
• 关键词: Animal Feed; Applied Genetics; Area; Binding; Binding Proteins; Capsid; Cells; Clathrin; Clathrin Heavy Chains; Complex; Cytoplasmic Structures; Data; Double Stranded RNA Virus; Edible Plants; Family; Food; Genetic; Genetic Complementation Test; Genome; Human; Infection; Lead; Learning; Literature; Livestock; Mammalian Orthoreovirus; Maps; Mediating; Methods; Microtubules; Modeling; Molecular; N-terminal; Nature; Oncolytic; Orbivirus; Play; Process; Proteins; Public Health; Publishing; Purpose; RNA Interference; RNA Viruses; Range; Recruitment Activity; Reoviridae Infections; Reovirus; Research Design; Role; Rotavirus; Screening procedure; Site; Structural Protein; Structure; Study models; Testing; Thinking; Viral; Viral Core Proteins; Viral Genome; Viral Pathogenesis; Viral Proteins; Virus; Virus Diseases; base; dimer; loss of function; member; mutant; nucleoside triphosphatase; particle; pathogen; positional cloning; protein protein interaction; prototype; research study; vector; viral RNA

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.

项目摘要：该项目的长期、广泛目标是了解细胞内、分子 哺乳动物正呼肠孤病毒（呼肠孤病毒）的机制，这是人类已知的致病原因 感染和建立的病毒发病机制研究模型，介导了复杂的、动态的 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 专注于病毒非结构蛋白 muNS，该蛋白已被证明在形成中发挥关键作用 特殊的细胞质结构，即“病毒工厂”，呼肠孤病毒复制和传播的许多步骤都在其中进行 组装发生。相关结构被同一分类的其他成员用于类似目的 多节段双链RNA病毒家族，例如人类病原体轮状病毒和 家畜病原体环状病毒。拟议的实验将尝试表征 muNS 蛋白的特征 决定其自我关联以及与细胞因子关联以诱导形成的能力 在没有其他呼肠孤病毒蛋白的情况下的工厂状内含物（目标 2）及其缔合能力 与其他病毒蛋白和潜在的 RNA 一起招募它们到工厂或类似工厂的内含物 （目标 1）。其他拟议的实验将尝试完善和应用遗传方法，包括 RNA 干扰和基于向量的反式互补，用于测试 muNS 和工厂的作用 呼肠孤病毒的生产性感染（目标 3）。在这些目标的框架内，其他病毒和细胞 建议对与 muNS 相关的成分进行研究，特别强调 病毒 sigmaNS 和 mu2 蛋白，它们也与确定病毒的特征或功能有关 工厂。与公共卫生的相关性：病毒基因组复制和颗粒的细胞内步骤 对于许多病毒的组装仍然知之甚少，一旦更好地理解，预计将呈现出 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。这 本提案重点关注的特定病毒，即哺乳动物呼肠孤病毒，是分类科的原型 其中包括人类、食用动物和食用植物的重要病原体，而且正在研究中 一种溶瘤剂。