MOLECULAR MECHANISMS OF NORWALK VIRUS GENOME EXPRESSION

Norwalk 病毒基因组表达的分子机制

• 批准号: 6534108
• 负责人: MICHELE E HARDY
• 金额: $ 12.98万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6534108
• 关键词: RNA virus; acute infectious nonbacterial gastroenteritis; cell free system; cell line; emerging infectious disease; gel mobility shift assay; gene expression; host organism interaction; immunoprecipitation; intermolecular interaction; intestines; laboratory rabbit; posttranslational modifications; protein biosynthesis; protein protein interaction; virus RNA; virus genetics; virus protein; virus replication

The long-term research objective of this laboratory is to understand in detail, the molecular mechanisms of Norwalk virus genome expression and replication. Norwalk virus (NV) is the prototype strain of non-cultivable human caliciviruses that are the most important cause of epidemic outbreaks of acute gastroenteritis in humans. NV is considered an emerging virus, as new data based on more sensitive molecular assays indicate that infections with these viruses are more widespread than originally recognized. NV is an exclusively human pathogen, but other caliciviruses infect a broad range of animals and cause persistent, chronic and sometimes lethal infections in their hosts. The ability of the caliciviruses to persist and the potential for emergence, both in prevalence and pathogenesis, exemplify the importance of understanding the replication strategies of the virus at the molecular level. This application proposed studies to understand the viral protein functions critical for replication of the NV genome, and interactions with cellular proteins that regulate these functions. The specific aims of this proposal are 1) To understand the mechanisms of synthesis and processing of the NV non-structural proteins. The details of polyprotein synthesis and post- translation processing will be studied by expression of NV RNA in intestinal cell-free extracts. 2) To understand the mechanisms of control of NV genome expression in intestinal cells. NV non-structural protein synthesis and processing will be investigated in transfected intestinal cells expressing the ORF1 polyprotein. 3) To understand the functional interactions of NV non-structural proteins. Specific interactions of non-structural proteins with RNA will be studied in intestinal cells expressing ORF1. NV and other caliciviruses are unique among animal viruses in structure and genome organization. Thus, detailed dissection of the functions of the NV non-structural proteins likely will lead to elucidation of new mechanisms of RNA virus genome expression.

本实验室的长期研究目标是详细了解诺瓦克病毒基因组表达和复制的分子机制。诺瓦克病毒（NV）是不可培养的人类杯状病毒的原型株，是人类急性肠胃炎流行暴发的最重要原因。NV被认为是一种新出现的病毒，因为基于更灵敏的分子测定的新数据表明，这些病毒的感染比最初认识到的更为广泛。NV是一种专门的人类病原体，但其他杯状病毒感染广泛的动物，并在其宿主中引起持续、慢性和有时致命的感染。杯状病毒在流行和发病机制方面持续存在的能力和出现的可能性，说明了在分子水平上理解病毒复制策略的重要性。该应用程序提出了研究，以了解对NV基因组复制至关重要的病毒蛋白功能，以及与调节这些功能的细胞蛋白的相互作用。本课题的具体目的是：1)了解NV非结构蛋白的合成和加工机制。通过在肠道无细胞提取物中表达NV RNA来研究多蛋白合成和翻译后加工的细节。2)了解NV基因组在肠细胞中的表达调控机制。将在表达ORF1多蛋白的转染肠细胞中研究NV非结构蛋白的合成和加工。3)了解NV非结构蛋白的功能相互作用。非结构蛋白与RNA的特异性相互作用将在表达ORF1的肠细胞中进行研究。NV和其他杯状病毒在结构和基因组组织方面在动物病毒中是独特的。因此，详细解剖NV非结构蛋白的功能可能会导致RNA病毒基因组表达的新机制的阐明。