Structure and Function of a Viral RNA-Dependent RNA Polymerase

病毒RNA依赖性RNA聚合酶的结构和功能

• 批准号: 9807800
• 负责人: C. Cheng Kao
• 金额: $ 30万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2002-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9807800&HistoricalAwards=false
• 关键词: Structure; Function; Viral; RNA; Dependent

Kao Viral RNA replication is a fundamental process in viral pathogenesis. This laboratory studies brome mosaic virus (BMV), a model (+)-strand RNA virus, in order to characterize the interactions between RNA-dependent RNA polymerase (RdRp) subunits and the mechanism of RNA synthesis. Three projects will be carried out simultaneously in this research. The first is designed to dissect the requirements for the interaction of the proteins found in the BMV replicase through chemical mutagenesis, selection for stable mutants that have lost protein-protein interaction in the two-hybrid assay, and sequencing the mutations. The second project is designed to elucidate the sequences that contribute to the initiation of (-)-strand viral RNA synthesis and to examine how these sequences affect steps in (-)-strand RNA synthesis. The third project examines genomic (+)-strand RNA synthesis by identifying the minimal functional promoter and constructing promoters with nucleotide substitutions, to identify RNA sequences that affect the various steps in viral RNA synthesis.Knowledge gained from the execution of this project will contribute to the basic understanding of how viruses replicate. The use of a plant virus, brome mosaic virus, as an experimental model, will directly increase the understanding of less well-studied viruses, some of which cause disease. The detailed understanding of the different steps in viral RNA replication could ultimately uncover targets for the design of highly specific antiviral compounds.

KAO病毒RNA复制是病毒致病过程中的一个基本过程。本实验室对一种模型(+)链RNA病毒--雀麦花叶病毒(BMV)进行了研究，以了解依赖RNA的RNA聚合酶(RdRp)亚基之间的相互作用和RNA合成的机制。这项研究将同时进行三个项目。第一项旨在通过化学诱变剖析BMV复制酶中发现的蛋白质相互作用的要求，选择在双杂交试验中失去蛋白质-蛋白质相互作用的稳定突变体，并对突变进行测序。第二个项目旨在阐明有助于启动(-)链病毒RNA合成的序列，并检查这些序列如何影响(-)链RNA合成的步骤。第三个项目通过确定最小的功能启动子和构建具有核苷酸替换的启动子来研究基因组(+)链RNA的合成，以确定影响病毒RNA合成的各个步骤的RNA序列。使用一种名为雀麦花叶病毒的植物病毒作为实验模型，将直接增加对研究较少的病毒的了解，其中一些病毒会导致疾病。对病毒RNA复制不同步骤的详细了解最终可能会发现设计高度特异的抗病毒化合物的目标。