ANALYSIS OF SPHERICAL VIRUS ASSEMBLY

球形病毒组装分析

• 批准号: 6099745
• 负责人: MARK j YOUNG
• 金额: --
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099745
• 关键词: Comovirus; X ray crystallography; gene expression; gene mutation; monoclonal antibody; nucleic acid sequence; protein sequence; ribonucleoproteins; site directed mutagenesis; structural biology; tissue /cell culture; virus RNA; virus assembly; virus protein

The assembly of spherical viruses is not well understood. The objective of this proposal is to examine the initial events in the assembly of a small icosahedral virus, cowpea chlorotic mottle virus (CCMV). The specific objectives of this proposal are to: (i) define and map the viral RNA and coat protein sequences responsible for initiating viral assembly; (ii) characterize the biochemical properties of the initiation complex for viral assembly; (iii) demonstrate both in vitro and in vivo that RNA packaging signals are essential for virus particle assembly and (iv) determine the role protein-protein and protein-RNA interactions contribute to the formation of the assembly initiation complex. The long-term goal of this research is to determine the precise mechanisms and regulation of viral assembly. An understanding of virion assembly may reveal ways to interfere with assembly, providing novel approaches to the control of viral diseases. CCMV provides an attractive system to unravel the assembly pathway because assembly questions can be addressed using the tools of molecular genetics, biochemistry, and structural biology. The assembly initiation complex will be characterized by three complementary approaches: (i) a biochemical approach which seeks to isolate the minimum ribonucleoprotein complex required to initiate viral assembly, (ii) a structural approach which takes advantage of the high resolution virus structure determined by X-ray crystallography and (iii) a molecular genetic approach in which mutations in the viral RNA are isolated which prevent assembly. Both in vitro and in vivo assembly assays will be used to characterize the initiation complex. Both the viral RNA sequences and the protein(s) constituting the origin of assembly will be examined.

球形病毒的组装尚不清楚。 目标 该建议的是检查组装中的初始事件 小型二十面体病毒，牛角绿化斑点病毒（CCMV）。 这 该提案的具体目标是：（i）定义和映射病毒 RNA和外套蛋白序列负责启动病毒组装； （ii）表征起始复合物的生化特性 病毒组装； （iii）在体外和体内都表明RNA 包装信号对于病毒颗粒组件至关重要，（iv） 确定蛋白质 - 蛋白质和蛋白-RNA相互作用的作用有助于 形成组装启动络合物。 长期目标 这项研究是确定的确切机制和调节 病毒组装。 对病毒装配的理解可能会揭示 干扰组装，提供了新颖的方法来控制 病毒疾病。 CCMV提供了一个吸引人的系统来揭示组件途径，因为 可以使用分子遗传学工具来解决集装问题， 生物化学和结构生物学。 装配启动综合体 将以三种互补方法为特征：（i）生化 试图隔离最小核糖核蛋白复合物的方法 启动病毒装配需要，（ii）一种结构方法 利用由X射线确定的高分辨率病毒结构 晶体学和（iii）一种分子遗传学方法，其中突变 在病毒RNA中，分离出来，以防止组装。 在体外和 体内组装测定将用于表征启动 复杂的。 病毒RNA序列和构成蛋白质的蛋白 将检查组装的起源。