RNA-dependent RNA Synthesis by the Hepatitis C Virus

丙型肝炎病毒依赖 RNA 的 RNA 合成

• 批准号: 8009848
• 负责人: Cheng C Kao
• 金额: $ 31.52万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8009848
• 关键词: Acquired Immunodeficiency Syndrome; Antiviral Agents; Binding; Biochemical; Biological; Biological Assay; Cells; Complex; DNA biosynthesis; Data; Development; Disease; Electron Microscopy; Genetic Transcription; Genome; Goals; Health; Hepatitis C; Hepatitis C virus; Human; Image; In Vitro; Knowledge; Laboratories; Lead; Learning; Left; Membrane; Methods; Modeling; Mutation; Peptide Hydrolases; Phase; Polymerase; Population; Principal Investigator; Property; Proteins; RNA; RNA Viruses; RNA chemical synthesis; RNA replication; RNA-Protein Interaction; Recombinants; Replicon; Research; Research Personnel; Resources; Staging; Structure; United States; Viral; Virus; Virus Replication; base; helicase; improved; in vitro activity; in vivo; liver transplantation; mutant; new technology; novel; programs; protein complex; protein protein interaction; repaired; replicase; vector; viral RNA

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) infects approximately 2% of the world's population and is the primary cause of liver transplants in the United States. Based on lessons learned from diseases such as AIDS, HCV RNA replication is a promising target for antiviral development. However, the replication of all viruses with plus-strand RNA genomes is poorly understood, especially at the biochemical level. The overall goal of the research in the Kao lab is to understand mechanism of RNA virus replication. The goal for this project is to build knowledge about the subunits of the HCV replication complex, with emphasis on protein-RNA, and protein-protein interactions. This is an extension of the past six years of research in the Kao lab, where a number of basic properties of the HCV polymerase and the HCV protease-helicase have been examined using biochemical, biophysical, and cell-based methods. The research can be partitioned into several related subaims that will: 1. Identify and validate the biological importance of the residues in the HCV RdRp that interacts with the substrate NTPs, the template RNA, and during different stages of HCV RNA synthesis. 2. Elucidate the interactions between the HCV RdRp and the nascent RNA and identify and characterize the nascent RNA exit channel. 3. Examine the protein-protein and protein-RNA interactions with other replicase- associated subunits of the HCV replicase and examine effects of the interactions on HCV replicase formation in cells. 4. Obtain images of the protein complexes using electron microscopy and reconstruct their structures. Results from this proposal will advance the understanding of the mechanism of viral RNA-dependent RNA synthesis for ALL positive-strand RNA viruses. The knowledge can also be used to compare the mechanisms of action of all template-dependent (both viral and cellular) polymerases.

描述（由申请人提供）：丙型肝炎病毒（HCV）感染了世界大约2％的人口，是美国肝移植的主要原因。基于从艾滋病等疾病中学到的经验教训，HCV RNA复制是抗病毒发育的有希望的目标。然而，对所有病毒的复制与链链RNA基因组的复制知之甚少，尤其是在生化水平上。 KAO实验室研究的总体目标是了解RNA病毒复制的机制。 该项目的目的是建立有关HCV复制复合物的亚基的知识，重点是蛋白-RNA和蛋白质 - 蛋白质相互作用。这是过去六年在KAO实验室进行研究的扩展，在该实验室中，已经使用了生物化学，生物物理和基于细胞的方法研究了HCV聚合酶和HCV蛋白酶丝酶的许多基本特性。这项研究可以分为几个相关的子iaims： 1。识别并验证与底物NTP，模板RNA以及HCV RNA合成的不同阶段相互作用的HCV RDRP中残基的生物学重要性。 2。阐明HCV RDRP与新生RNA之间的相互作用，并识别和表征新生的RNA出口通道。 3。检查蛋白质 - 蛋白质蛋白和蛋白-RNA与HCV复制酶的其他相关亚基的相互作用，并检查相互作用对细胞中HCV复制酶形成的影响。 4。使用电子显微镜获得蛋白质复合物的图像并重建其结构。 该提案的结果将提高对所有阳性RNA病毒病毒RNA依赖性RNA合成机制的理解。知识还可以用来比较所有模板依赖性（病毒和细胞）聚合酶的作用机理。