CELL BIOLOGY OF VIRUS ENTRY

病毒进入的细胞生物学

• 批准号: 2060733
• 负责人: ARI H HELENIUS
• 金额: $ 30.11万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-01-01 至 1996-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2060733
• 关键词: Alphavirus; HeLa cells; Orthomyxoviridae; Semliki Forest virus; Vesiculovirus; acids; amantadine; antiviral agents; antiviral antibody; cell fusion; chloroquine; cholesterol; chromosome aberrations; conformation; electron microscopy; genetic manipulation; genetic transcription; genetic translation; glycoproteins; host organism interaction; immunocytochemistry; influenza; laboratory mouse; laboratory rabbit; macromolecule; membrane activity; membrane fusion; membrane lipids; microinjections; microorganism genetics; microorganism hemagglutinin; molecular pathology; monoclonal antibody; nucleocapsid; oligonucleotides; phagocytosis; poliovirus; radiotracer; receptor mediated endocytosis; temperature sensitive mutant; tissue /cell culture; virus DNA; virus antigen; virus cytopathogenic effect; virus envelope; virus genetics; virus infection mechanism; virus protein; virus replication; viruslike particle

We are studying the mechanism whereby viruses enter their host cells and initiate replication. The virus systems to be studied are Semliki Forest virus (SFV) and Influenza A virus, which have proven relevant model systems for the general problems of early virus-cell interactions. The overall aim is to define on molecular and cellular terms the following steps in entry: internalization of the virus particles by endocytosis, penetration from endosomes by membrane fusion, uncoating of the genome, and transport to the cytoplasmic or nuclear location of replication and transcription. We hope to obtain a deeper understanding of the principles underlying viral pathogenicity and cell tropism as well as of basic biological phenomena such as membrane fusion, nuclear targeting and the interaction between cellular compartments. The specific aspects include a more detailed definition of endocytic organelles where acid-triggered fusion of the viruses takes place. The conformational changes in the viral fusion proteins acid-induced, activity will be characterized in detail with emphasis on the sterol dependence of SFV (now recognized as a common property among viruses) and the interaction between the fusion factors and the fusing membranes. The mechanism of action of the trimeric hemagglutinin, the fusion factor of Influenza virus, will be studied using hybrid molecules made up of trimers consisting of different subunits. The fate of the nucleocapsids after penetration will be followed by biochemical and immunochemical methods. We will, moreover, try to resolve the signals which induce uncoating of the genome and analyze the molecular mechanism by which uncoating takes place, the transport of the genome to the SFV-specific cytopathic vacuols, and Influenza to the nucleus, will be investigated. A variety of methods ranging from immunocytochemistry on frozen sections to bulk delivery of anti-sense oligonucleotides will be used.

我们正在研究病毒进入其宿主的机制 细胞并启动复制。 要研究的病毒系统是 Semliki森林病毒（SFV）和流感病毒，具有 已验证的相关模型系统，用于早期的一般问题 病毒细胞相互作用。 总体目的是定义分子 和蜂窝术语的条目以下步骤： 内吞作用的病毒颗粒，从内体渗透 膜融合，基因组的脱膜和运输到 复制和转录的细胞质或核位置。 我们希望对原则有更深入的了解 潜在的病毒致病性和细胞托管以及基本 生物学现象，例如膜融合，核靶标 以及细胞隔室之间的相互作用。 具体方面包括更详细的定义 酸触发病毒融合的内吞细胞器 发生。 病毒融合的构象变化 蛋白质酸诱导的活性将详细表征 强调SFV的固醇依赖性（现在被公认为是 病毒之间的共同特性）以及 融合因子和融合膜。 机制 三聚体血凝素的作用，融合因子的融合因子 流感病毒将使用由由 由不同亚基组成的三聚体。 命运 穿透后的核蛋白会随后发生生化 和免疫化学方法。 此外，我们将尝试解决 诱导基因组脱膜并分析的信号 发生脱衣的分子机制， 将基因组传输到SFV特异性细胞疗法真空吸尘器， 将研究对核的流感。 各种各样 从冷冻切片的免疫细胞化学范围 将使用反义寡核苷酸的批量输送。