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: 7383968
• 负责人: Cheng C Kao
• 金额: $ 28.77万
• 依托单位: TEXAS A&M 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 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7383968
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Antiviral Agents; Biochemical; Biological; Cells; Complement component C1s; Complex; DNA biosynthesis; Development; Disease; Electron Microscopy; Endopeptidases; Genetic Transcription; Genome; Goals; Health; Hepatitis C; Hepatitis C virus; Human; Image; In Vitro; Knowledge; Laboratories; Learning; Membrane; Methods; Modeling; Numbers; Peptide Hydrolases; Phase; Polymerase; Population; Property; Proteins; RNA; RNA Viruses; RNA chemical synthesis; RNA replication; RNA-Protein Interaction; Recombinants; Research; Staging; Structure; United States; Viral; Viral Proteins; Virus; Virus Replication; base; helicase; liver transplantation; novel; protein protein interaction; replicase; 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）感染约2%的世界人口，是美国肝移植的主要原因。基于从艾滋病等疾病中吸取的教训，HCV RNA复制是抗病毒药物开发的一个有希望的靶点。然而，对所有具有正链RNA基因组的病毒的复制知之甚少，特别是在生物化学水平上。Kao实验室研究的总体目标是了解RNA病毒复制的机制。 该项目的目标是建立有关HCV复制复合体亚基的知识，重点是蛋白质-RNA和蛋白质-蛋白质相互作用。这是Kao实验室过去六年研究的延伸，在那里，HCV聚合酶和HCV蛋白酶解旋酶的一些基本特性已经使用生物化学，生物物理和基于细胞的方法进行了检查。这项研究可以分为几个相关的子目标，这些子目标将： 1.鉴定并验证HCV RdRp中与底物NTP、模板RNA相互作用的残基以及HCV RNA合成不同阶段的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，并鉴定和表征新生RNA的出口通道。 3.检查蛋白质-蛋白质和蛋白质-RNA与HCV复制酶的其他复制酶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。 这一提议的结果将促进对ALL正链RNA病毒RNA依赖性RNA合成机制的理解。这些知识也可用于比较所有模板依赖性（病毒和细胞）聚合酶的作用机制。