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 该提案的目的是审查大会的最初事件 小二十面体病毒，豇豆褪绿斑驳病毒（CCMV）。 这 该提案的具体目标是： (i) 定义并绘制病毒式传播 负责启动病毒组装的RNA和外壳蛋白序列； (ii) 表征起始复合物的生化特性 病毒组装； (iii) 在体外和体内证明 RNA 包装信号对于病毒颗粒组装至关重要，并且 (iv) 确定蛋白质-蛋白质和蛋白质-RNA 相互作用的作用 组装起始复合物的形成。 长期目标 这项研究的目的是确定精确的机制和调节 病毒组装。 对病毒粒子组装的理解可能会揭示以下方法： 干扰组装，提供了控制的新方法 病毒性疾病。 CCMV 提供了一个有吸引力的系统来解开组装途径，因为 组装问题可以使用分子遗传学工具来解决， 生物化学和结构生物学。 组装起始复合体 其特点是三种互补的方法：(i) 生化方法 寻求分离最小核糖核蛋白复合物的方法 启动病毒组装所需的，（ii）一种结构方法 利用 X 射线确定的高分辨率病毒结构 晶体学和（iii）分子遗传学方法，其中突变 病毒RNA中的RNA被分离出来，防止组装。 无论是体外还是 体内组装测定将用于表征起始 复杂的。 病毒RNA序列和构成病毒的蛋白质 将检查装配的起源。