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：
7385110
负责人：
Olve Breien Peersen
金额：
$ 23.38万
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 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7385110
关键词：
Active Sites Acute Hepatitis Affect Biochemical Common Cold Complex Data Dimerization Disease Endopeptidases Endoplasmic Reticulum Enzymes Family Family Picornaviridae Fingers Generations Heart Diseases Hepatitis A Virus Human poliovirus Hydrogen Bonding In Vitro Life Cycle Stages Membrane Methods Molecular Molecular Conformation Mutation Nucleotides Paralysed Peptide Hydrolases Peptides Pliability Polioviruses Polymerase Polyproteins Positioning Attribute Process Protein Precursors Proteins RNA RNA Binding RNA Viruses RNA chemical synthesis RNA replication RNA-Directed RNA Polymerase Range Replication-Associated Process Research Personnel Resolution Rhinovirus Role Staging Structure Surface Thermodynamics Vesicle Viral Virus base cost in vivo insight member novel pathogen poliovirus polymerase 3Dpol programs three dimensional structure tripolyphosphate 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 common life cycle where their 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 the synthesis of all viral RNA. Like all picornaviral proteins, the polymerase is generated by proteolytic cleavage of a single large viral polyprotein. There is mounting evidence in several picornaviruses that the polymerase and its immediate precursors are directly responsible for the assembly of these replication centers. The 3Dpol polymerase of poliovirus, the best studied of the picornaviruses, has been shown to assemble into large sheet structures along a protein-protein interface that was initially identified in a partial crystal structure of 3Dpol. We have solved the complete crystal structure of 3Dpol at 2.0 A resolution and discovered that the enzyme requires a free N-terminus to properly fold the active site, providing a molecular basis for the processing dependent activation of the polymerase. We are continuing our structural studies of the poliovirus proteins by further characterizing the conformational flexibility of 3Dpol and expanding into determining the role of protein-protein interfaces in the assembly of 3CDpro and other precursor proteins. The results will yield fundamental insights into the replication of this important group of pathogens.

描述（由申请人提供）：小核糖核酸病毒是一类小型正链单链RNA病毒，可引起多种疾病，每年造成的损失高达数亿美元。其成员包括急性甲型肝炎病毒、引起心脏病的柯萨奇 B3 病毒、导致一半以上普通感冒发生的鼻病毒以及麻痹性脊髓灰质炎病毒。这些病毒具有共同的生命周期，其中它们的RNA复制和病毒组装发生在源自内质网的囊泡表面组装的大膜锚定复制复合物中。复制过程由病毒编码的 RNA 依赖性 RNA 聚合酶（3Dpol 蛋白）驱动，该酶负责所有病毒 RNA 的合成。与所有小核糖核酸病毒蛋白一样，聚合酶是通过单个大病毒多蛋白的蛋白水解裂解产生的。在几种小核糖核酸病毒中，越来越多的证据表明聚合酶及其直接前体直接负责这些复制中心的组装。脊髓灰质炎病毒的 3Dpol 聚合酶是研究最深入的小核糖核酸病毒，已被证明可以沿着蛋白质-蛋白质界面组装成大的片状结构，该界面最初是在 3Dpol 的部分晶体结构中发现的。我们以 2.0 A 分辨率解析了 3Dpol 的完整晶体结构，并发现该酶需要游离的 N 末端才能正确折叠活性位点，为聚合酶的加工依赖性激活提供了分子基础。我们正在继续对脊髓灰质炎病毒蛋白进行结构研究，进一步表征 3Dpol 的构象灵活性，并扩展到确定蛋白质-蛋白质界面在 3CDpro 和其他前体蛋白组装中的作用。研究结果将为这一组重要病原体的复制提供基本见解。