Gene Functions and Molecular Genetics of RNA Viruses

RNA病毒的基因功能和分子遗传学

• 批准号: 7531819
• 负责人: PAUL G. AHLQUIST
• 金额: $ 42.15万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7531819
• 关键词: Affinity; Alphavirus; Area; Bacteriophages; Binding; Biochemical; Biological Models; Biotechnology; Brome mosaic virus; Capsid; Cataloging; Catalogs; Cell physiology; Cells; Cellular biology; Complex; Cytopathology; Cytoplasm; Disease; Double Stranded RNA Virus; Double-Stranded RNA; Electron Microscope; Elements; Gene Expression; Genes; Genetic; Integration Host Factors; Libraries; Link; Maps; Medicine; Membrane; Membrane Lipids; Membrane Microdomains; Membrane Proteins; Modeling; Molecular Genetics; Morphogenesis; Neck; Normal Cell; Organelles; Peripheral; Plants; Process; Proteins; RNA; RNA Viruses; RNA chemical synthesis; RNA replication; Recruitment Activity; Regulation; Replication-Associated Process; Research; Resources; Retroviridae; Role; Signal Transduction; Staging; Structure; System; Testing; Three-Dimensional Imaging; Translations; Vesicle; Viral Proteins; Virion; Virus; Virus Diseases; Virus Replication; Yeasts; electron density; gene function; genome wide association study; genome-wide; genome-wide analysis; helicase; improved; in vivo; member; novel; nucleoside triphosphatase; programs; research study; tomography; viral RNA; virus host interaction

The long term objectives of this research program are to define the mechanisms of fundamental replication processes, virus-host interactions, and cytopathologies common to many important (+) strand RNA viruses, and to use these results to improve virus control and beneficial uses of viruses and their components in biotechnology and medicine. New directions on the functions and interactions of viral proteins, RNAs and host factors in RNA replication and gene expression will be pursued using brome mosaic virus (BMV), a representative member of the alphavirus-like superfamily and a productive, advanced model system. Valuable new opportunities in these areas follow from our recent findings that BMV RNA replication occurs in virus-induced mini-organelles (spherules) whose structure, assembly and function show parallels with replicative cores of retroviruses and dsRNA viruses; from our advancing definition of the functions and interactions of BMV replication factors 1a, 2ap0' and cis-acting RNA signals; and from our use of the rare ability of BMV to replicate in yeast to conduct the first genome-wide analysis of host gene functions in virus replication. Building on these, we will use BMV replication in natural plant host cells and yeast with synergistic genetic, biochemical and cell biology approaches, to analyze virus replication and virus-host interactions. Among other steps, we will use electron microscope tomography and related approaches to define the three dimensional in vivo ultrastructure of RNA replication complexes arrested at consecutive stages, to resolve crucial issues about their organization, morphogenesis and function. The mechanisms by which BMV 1a protein alone induces these membrane-enveloped spherule replication compartments will be determined, including the roles of multiple 1a-1a interactions and 1a interactions with membranes and lipid rafts. We will determine how 1a interaction with specific cis-acting viral RNA signals inhibits translation and recruits RNAs to replication, and if, as our results suggest, the 1a NTPase/helicase domain parallels dsRNA phage NTPases in translocating viral RNA replication templates into pre-formed replication compartments. We will analyze the contributions of selected host genes implicated in RNA replication, and use novel genetic approaches to perform genome-wide cataloging of the contributions of essential host genes to RNA replication. The results will advance understanding and control of widespread viral diseases, and understanding of normal cell functions relevant to other disease processes.

该研究计划的长期目标是定义基本复制的机制 许多重要（+）链RNA病毒常见的过程，病毒宿主相互作用和细胞病理学， 并使用这些结果来改善病毒控制和对病毒及其成分的有益用途 生物技术和医学。关于病毒蛋白，RNA和 RNA复制和基因表达中的宿主因子将使用Brome Mosaic病毒（BMV），A 类似α病毒的超家族和高级模型系统的代表成员。 从我们最近的发现，BMV RNA复制发生在这些领域的宝贵新机会 在病毒诱导的迷你器官（球形）中，其结构，组装和功能表明与 逆转录病毒和DSRNA病毒的复制核心；根据我们对功能的定义的定义 BMV复制因子1A，2AP0'和顺式作用RNA信号的相互作用；从我们使用稀有的 BMV在酵母中复制的能力对病毒中宿主基因功能进行首次全基因组分析 复制。在这些基础上，我们将在天然植物宿主细胞和酵母中使用BMV复制 协同遗传，生化和细胞生物学方法分析病毒复制和病毒宿主 互动。除其他步骤外，我们将使用电子显微镜断层扫描和相关方法 定义在连续捕获的RNA复制复制复合物的体内三维超微结构 阶段，解决有关其组织，形态发生和功能的关键问题。机制 BMV 1A蛋白单独诱导这些膜开发的球形复制室将 确定，包括多个1A-1A相互作用和1A相互作用与膜和1A相互作用的作用 脂筏。我们将确定与特定顺式作用病毒RNA信号的1A相互作用如何抑制翻译 并招募RNA进行复制，并且如我们的结果所表明的那样，1A NTPase/Helicase结构域的相似之处 DSRNA噬菌体NTPases在将病毒RNA复制模板转移到预先形成的复制中 车厢。我们将分析与RNA复制有关的选定宿主基因的贡献，并 使用新型的遗传方法来执行本质宿主的贡献的全基因组分类 基因复制。结果将提高对广泛病毒疾病的理解和控制， 并了解与其他疾病过程相关的正常细胞功能。