Assembly of Picornaviral Replication Complexes

小核糖核酸病毒复制复合物的组装

基本信息

批准号：
8277241
负责人：
Olve Breien Peersen
金额：
$ 34.11万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8277241
关键词：
ATP phosphohydrolase Active Sites Acute Hepatitis Affect Antiviral Agents Back Binding Biochemical Biochemistry Catalysis Cell membrane Cells Cellular Membrane Common Cold Complex Coxsackie Viruses DNA Sequence Rearrangement DNA-Directed RNA Polymerase Data Detergents Development Disease Endoplasmic Reticulum Engineering Family Family Picornaviridae Funding Future Genome Genomics Goals Heart Diseases Hepatitis A Virus Human poliovirus In Vitro Individual Length Life Cycle Stages Maps Membrane Membrane Proteins Membrane Structure and Function Methods Molecular Motor Mutation Nucleotides Paralysed Peptide Hydrolases Play Polioviruses Polymerase Polyproteins Positioning Attribute Protein Precursors Proteins Proteolytic Processing RNA RNA Viruses RNA chemical synthesis RNA replication RNA-Binding Proteins RNA-Directed RNA Polymerase Reaction Replication-Associated Process Research Research Project Grants Resolution Rhinovirus Site Staging Structure Surface System Testing Translations Update Vaccines Vesicle Viral Viral Genome Virus Virus Diseases Virus Replication base cost helicase in vitro activity in vivo member mutant nanodisk porin protein activation protein function public health relevance reconstitution research study viral RNA

项目摘要

DESCRIPTION (provided by applicant): The picornaviruses are a family of small positive sense single stranded RNA viruses that cause a wide range of diseases at an annual cost well into the hundreds of million dollars. Members include acute hepatitis A virus, the heart disease causing coxsackie B3 virus, rhinoviruses that cause more than half the occurrences of the common cold, and the paralyzing poliovirus. These viruses share a life cycle where RNA replication and viral assembly occurs in large membrane anchored replication complexes assembled on the surfaces of vesicles derived from the endoplasmic reticulum. The replication process is driven by a virally encoded RNA dependent RNA polymerase, the 3Dpol protein, that is responsible for the synthesis of all viral RNA. This research project is focused on the structure and assembly of viral replication centers, where we use poliovirus and coxsackievirus as our main experimental systems. We have previously solved the crystal structures of the 3Dpol proteins from both these viruses and elucidated the molecular mechanism behind the proteolytic activation of these proteins upon cleavage from the viral 3CDpro precursor protein. We are continuing our studies of picornaviral replication proteins by focusing on the structural changes associated with the formation of the 3Dpol elongation complex and on understanding how mutations in the polymerase affect viral RNA replication rate and fidelity both in vitro and in vivo. A new aspect of the project is focused on the structure and function of the membrane associated viral 2C and 2BC proteins that are responsible for host cell membrane rearrangements resulting in the formation of the vesicles upon which the viral replication complexes assemble. PUBLIC HEALTH RELEVANCE: Poliovirus is a member of a large family of viruses containing a specific RNA dependent RNA polymerase protein that is responsible for replicating the viral genome. The focus of this research project is to understand how this protein functions during virus replication and find ways to interfere with its function, which can open the door to the development of antiviral drugs.

描述（由申请人提供）：Picornaviruses是一个小型积极的单一链RNA病毒的家族，每年以每年成本良好的成本造成多种疾病。成员包括急性肝炎病毒，心脏病，引起Coxsackie B3病毒，鼻病毒导致普通感冒的一半以上以及瘫痪的脊髓灰质炎病毒。这些病毒具有生命周期，其中RNA复制和病毒组装发生在大膜锚定复制复合物中，这些复制复制复合物组装在源自内质网的囊泡表面上。复制过程是由病毒编码的RNA依赖性RNA聚合酶，3DPOL蛋白的驱动，该蛋白是所有病毒RNA的合成。该研究项目的重点是病毒复制中心的结构和组装，我们将poliovirus和coxsackievivirus用作我们的主要实验系统。我们先前已经从这些病毒中求解了3DPol蛋白的晶体结构，并阐明了这些蛋白质在从病毒3CDPRO前体蛋白上切割后这些蛋白质的蛋白水解活化背后的分子机制。我们通过关注与3DPol伸长复合物的形成相关的结构变化，并了解聚合酶中的突变如何影响病毒RNA复制率和体内的忠诚度，从而继续对Picornaviral复制蛋白进行研究。该项目的一个新方面集中在膜相关病毒2C和2BC蛋白的结构和功能上，该蛋白负责宿主细胞膜重排，从而形成病毒复制复合物的囊泡。公共卫生相关性：脊髓灰质炎病毒是含有特定RNA依赖RNA聚合酶蛋白的大型病毒家族的成员，该蛋白负责复制病毒基因组。该研究项目的重点是了解该蛋白在病毒复制过程中的功能，并找到干扰其功能的方法，这可以为抗病毒药的发展打开大门。