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: 8204883
• 负责人: Cheng C Kao
• 金额: $ 33.29万
• 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 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204883
• 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) 感染世界上大约 2% 的人口，是美国肝移植的主要原因。根据艾滋病等疾病的经验教训，HCV RNA 复制是抗病毒开发的一个有希望的目标。然而，人们对所有具有正链RNA基因组的病毒的复制知之甚少，特别是在生化水平上。花王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。这是花王实验室过去六年研究的延伸，其中使用生物化学、生物物理和基于细胞的方法检查了 HCV 聚合酶和 HCV 蛋白酶解旋酶的许多基本特性。该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 相互作用以及在 HCV RNA 合成的不同阶段相互作用的残基的生物学重要性。 2. 阐明HCV RdRp 和新生RNA 之间的相互作用，并鉴定和表征新生RNA 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。 该提案的结果将促进对所有正链 RNA 病毒的病毒 RNA 依赖性 RNA 合成机制的理解。这些知识还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。

项目成果

Crystal structure of RIG-I C-terminal domain bound to blunt-ended double-strand RNA without 5' triphosphate.

• DOI: 10.1093/nar/gkq974
• 发表时间: 2011-03
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Lu C;Ranjith-Kumar CT;Hao L;Kao CC;Li P
• 通讯作者: Li P

The structural basis of 5' triphosphate double-stranded RNA recognition by RIG-I C-terminal domain.

• DOI: 10.1016/j.str.2010.05.007
• 发表时间: 2010-08-11
• 期刊: Structure (London, England : 1993)
• 作者: Lu C;Xu H;Ranjith-Kumar CT;Brooks MT;Hou TY;Hu F;Herr AB;Strong RK;Kao CC;Li P
• 通讯作者: Li P

Identification and functional characterization of the nascent RNA contacting residues of the hepatitis C virus RNA-dependent RNA polymerase.

• DOI: 10.1261/rna.031914.111
• 发表时间: 2012-08
• 期刊: RNA
• 影响因子: 4.5
• 作者: Robert C Vaughan;B. Fan;J. You;C. Kao

The hepatitis C virus core protein can modulate RNA-dependent RNA synthesis by the 2a polymerase.

丙型肝炎病毒核心蛋白可以通过 2a 聚合酶调节 RNA 依赖性 RNA 合成。

• DOI: 10.1016/j.virusres.2014.05.017
• 发表时间: 2014
• 期刊: Virus research
• 影响因子: 5
• 作者: Wen,Y;ChengKao,C
• 通讯作者: ChengKao,C

Mutations in the hepatitis C virus polymerase that increase RNA binding can confer resistance to cyclosporine A.

• DOI: 10.1002/hep.22987
• 发表时间: 2009-07
• 期刊: HEPATOLOGY
• 影响因子: 13.5
• 作者: Liu, Zhe;Robida, John M.;Chinnaswamy, Sreedhar;Yi, Guanghui;Robotham, Jason M.;Nelson, Heather B.;Irsigler, Andre;Kao, C. Cheng;Tang, Hengli
• 通讯作者: Tang, Hengli