Hepatitis C virus genome structure: dynamic roles in replication and infectivity

丙型肝炎病毒基因组结构：复制和感染性中的动态作用

基本信息

批准号：
9980781
负责人：
Brett D. Lindenbach
金额：
$ 55.3万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9980781
关键词：
Animals Antiviral Agents Architecture Behavior Binding Biochemical Biochemistry Biological Models Cells Communities Complement Complex Crystallography Dengue Virus Disease Elements Enzymatic Biochemistry Event Funding Future Genetic Genome Health Hepatitis C virus Human In Vitro Individual Infection Integration Host Factors Knowledge Light Malignant neoplasm of liver Maps Methods Molecular Monitor Nonstructural Protein Phase Play Population Process Proteins Public Health RNA RNA Binding RNA Biochemistry RNA Viruses RNA replication RNA-Protein Interaction Reagent Regulation Replication Initiation Research Research Project Grants Role Rubber Site Specificity Structure Technology Testing Therapeutic Translating United States Untranslated RNA Viral Viral Genome Viral Nonstructural Proteins Viral Physiology Viral Proteins Virus Virus Assembly Virus Replication Work chronic liver disease combat crosslink design genetic approach genomic RNA helicase improved in vivo innovation insight interdisciplinary approach novel strategies protein complex protein function recruit replicase structural biology success tool viral RNA virology virus genetics

项目摘要

Project Summary Hepatitis C virus (HCV) infects a large population within the United States (~2%), where it remains a major cause of chronic liver disease and cancer. While potent antiviral drugs have recently become available, long-term success in combating HCV-related disease requires a clear understanding of viral replication mechanisms. Furthermore, HCV provides an essential model system for understanding the replication of many related positive-strand RNA viruses, including dengue virus and other unmet threats to human and animal health. To date, most work on HCV replication has focused on activities of the viral nonstructural (NS) proteins or the role of host factors in this process. Very little has been done on role of the HCV RNA genome in replication or on the functional interactions between the viral genome and NS proteins. This is a critical gap, since the interaction between HCV RNA and viral proteins is literally where the “rubber hits the road”, and the key events in viral replication occur. We will leverage our recent success in defining molecular networks among the NS proteins, and in characterizing RNA and RNA-protein complexes, and we will exploit new approaches for monitoring RNA interactions in vivo in order to achieve the following three aims: (A) Determine the functional RNA structures within the HCV genome; (B) Define the molecular interactions between NS proteins and the RNA genome, establishing their interaction sites and functional relevance during sequential phases of the viral lifecycle; (C) Define the structural features and functional attributes of the Replication Pre-initiation Complexes (RPIC), which initiates the entire process of virus propagation. These objectives will be accomplished by integrating innovative approaches for studying viral genetics, RNA crosslinking and sequencing, enzymology and crystallography, and capitalizing on our respective strengths in virology and RNA biochemistry. By focusing on the molecular interplay between genomic RNA and NS proteins, the work is conceptually innovative. In developing the tools needed to advance this research, the project is also technologically innovative, resulting in methods and reagents that will be of widespread utility to the RNA virus research community.

项目摘要丙型肝炎病毒（HCV）感染了美国大量人口（约2％）慢性肝病和癌症的主要原因。虽然最近有效的抗病毒药已成为可用的长期成功在对抗HCV相关疾病方面需要清楚地了解病毒复制机制。此外，HCV提供了理解的基本模型系统许多相关阳性RNA病毒的复制，包括登革热病毒和其他未经对人类和动物健康的威胁。迄今为止，大多数关于HCV复制的工作都集中在活动上病毒非结构（NS）蛋白质或宿主因素在此过程中的作用。几乎没有对HCV RNA基因组在复制或病毒之间的功能相互作用中所做的作用基因组和NS蛋白。这是一个关键的差距，因为HCV RNA与病毒之间的相互作用蛋白质实际上是“橡胶撞到道路”的地方，并且发生病毒复制中的关键事件。我们将利用我们最近在定义NS蛋白之间的分子网络方面的成功，在表征RNA和RNA-蛋白质复合物，我们将利用新方法进行监测为了实现以下三个目的，体内RNA相互作用：（a）确定功能 HCV基因组中的RNA结构；（b）定义NS蛋白之间的分子相互作用和RNA基因组，在顺序期间建立其相互作用位点和功能相关性病毒生命周期的阶段；（c）定义结构特征和功能属性复制预生效复合物（RPIC），启动了整个病毒传播过程。这些目标将通过整合研究病毒的创新方法来实现遗传学，RNA交联和测序，酶学和晶体学，并利用我们的病毒学和RNA生物化学方面的优势。通过关注分子相互作用在基因组RNA和NS蛋白之间，这项工作在概念上是创新的。在开发工具时为了推进这项研究，该项目在技术上也是创新的，导致方法以及对RNA病毒研究界的宽度实用性的试剂。