The Chemical Basis of Viral Assembly and Disassembly

MCB 9723752 Mark Young（蒙大拿州立大学）病毒组装和复制的化学基础 摘要 1.技术 本研究使用豇豆褪绿斑驳病毒（CCMV），一个成员的溴病毒组的Bromoviridea家庭，作为一个独特的模型系统，以确定详细的化学作用蛋白质-蛋白质和蛋白质-RNA相互作用的组装，成熟和拆卸过程中发挥的病毒。 CCMV是一种二十面体病毒，由四组分正链RNA基因组和衣壳组成，衣壳含有180个相同的外壳蛋白亚基，每个亚基的分子量为19.8KD。本提案的具体目标是：（1）确定关键的病毒RNA-外壳蛋白和外壳蛋白-外壳相互作用，这些相互作用作为调节病毒粒子结构和稳定性的分子决定因素;（2）确定N-外壳蛋白的末端进行从病毒体内部到病毒体外部的结构转变，以形成病毒体通过的通道RNA在分解过程中实验方法结合了分子和结构工具。产生具有改变的病毒体结构和稳定性的基因突变，并检查其对体外和体内病毒体组装和拆卸的影响，以便可以鉴定关键组分。 这项研究的结果将对广泛的动物、植物和昆虫病毒产生重大影响，并为预防病毒感染的策略提供基础。 2.非技术 了解蛋白质-蛋白质和蛋白质-核酸相互作用的化学基础是现代生物学的主要目标之一。 病毒粒子的组装、成熟和拆卸特别适合于确定这些相互作用的基本原理。 本研究的目的是确定病毒组装和拆卸过程中发生的详细分子相互作用如何影响和调节病毒的感染性。球形豇豆褪绿斑驳病毒是本研究项目的主题。生物化学和结构的方法来分析遗传突变体被用来确定的关键组成部分。这些发现将有助于了解病毒如何形成和破坏的基本知识，并为设计预防植物和动物病毒感染的策略提供重要信息。

MCB 9723752 Mark Young (Montana State University) The Chemical Basis of Viral Assembly and Disassembly ABSTRACT 1. Technical This study uses the cowpea chlorotic mottle virus (CCMV), a member of the bromovirus group of the Bromoviridea family, as a unique model system to determine the detailed chemical role protein-protein and protein-RNA interactions play in the assembly, maturation, and disassembly processes of a virus. An icosahedral virus, CCMV is constituted by a four component positive sense stranded RNA genome and capsids containing 180 identical coat protein subunits, each of 19.8KD. The specific objectives of this proposal are to: (1) Determine the critical viral RNA-coat protein and coat protein-coat interactions that act as molecular determinants regulating virion architecture and stability; and (2) Determine the ability of the N-terminus of the coat protein to undergo a structural transition from the interior of the virion to the exterior of the virion in order to form a channel for the passage of virion RNA during disassembly. The experimental approach combines molecular and structural tools. Genetic mutations with altered virion architecture and stability are generated and examined for their effect on virion assembly and disassembly in vitro and in vivo, so that critical components can be identified. The results of this research will have significant implications for a broad range of animal, plant, and insect viruses and should provide the basis for strategies for preventing viral infections. 2. Non-technical Understanding the chemical basis of protein-protein and protein-nucleic acid interactions is one of the major goals of modern biology. Virion assembly, maturation, and disassembly is particularly suitable for determining the underlying principles of these interactions. The objective of this study is to determine how the detailed molecular interactions occurring during virus assembly and disassembly influence and regulate virus infectivity. The spherical , cowpea chlorotic mottle virus is the subject of this research project. Biochemical and structural approaches to the analysis of genetic mutants are used to identify the critical components. The findings should contribute to the basic knowledge of how viruses are formed and disrupted, and provide essential information for the design of strategies to prevent viral infection to plants and animals.