MOLECULAR MECHANISMS OF NORWALK VIRUS GENOME EXPRESSION

Norwalk 病毒基因组表达的分子机制

• 批准号: 6373884
• 负责人: MICHELE E HARDY
• 金额: $ 13.85万
• 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 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6373884
• 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。诺沃克病毒(Norwalk Virus，NV)是不可培养的人类杯状病毒的原型株，是人类急性胃肠炎暴发流行的最重要原因。NV被认为是一种新兴病毒，因为基于更敏感的分子分析的新数据表明，这些病毒的感染比最初认识到的更广泛。新城疫病毒只是一种人类病原体，但其他杯状病毒可感染多种动物，并在宿主中造成持续性、慢性、有时甚至致命的感染。杯状病毒的持续能力和出现的可能性，无论是在流行还是在发病机制上，都证明了在分子水平上了解病毒复制策略的重要性。这项申请提出了研究，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。这项建议的具体目的是：1)了解NV非结构蛋白的合成和加工机制。多蛋白合成和翻译后处理的细节将通过NV RNA在肠道无细胞提取物中的表达来研究。2)了解NV基因组在肠道细胞中表达的调控机制。NV非结构蛋白的合成和加工将在表达ORF1多蛋白的转基因肠道细胞中进行研究。3)了解NV非结构蛋白的功能相互作用。非结构蛋白与RNA的特定相互作用将在表达ORF1的肠道细胞中进行研究。新城疫病毒和其他杯状病毒在结构和基因组组织上是动物病毒中独一无二的。因此，对NV非结构蛋白功能的详细剖析可能有助于阐明RNA病毒基因组表达的新机制。