ANALYSIS OF SPHERICAL VIRUS ASSEMBLY

球状病毒的组装还不是很清楚。目标是 这项提议的目的是审查在一个 小二十面体病毒、豌豆褪绿斑病毒(CCMV)。这个 这项提案的具体目标是：(I)界定和绘制病毒 负责启动病毒组装的RNA和外壳蛋白序列； (Ii)表征引发络合物的生化性质 病毒组装；(Iii)在体外和体内证明RNA 包装信号对病毒颗粒组装和(Iv)至关重要 确定蛋白质-蛋白质和蛋白质-RNA相互作用的作用 与组装引发复合体的形成有关。长期目标 这项研究的目的是确定其确切的机制和调节 病毒组装。对病毒粒子组装的了解可能会揭示 干扰装配，提供新的方法来控制 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的工具来解决， 生物化学和结构生物学。组装引发复合体 将通过三种相辅相成的方法来描述：(一)生化 寻求分离最小核糖核蛋白复合体的方法 启动病毒组装所需的，(Ii)结构方法，该方法 利用X射线确定的高分辨率病毒结构 结晶学和(Iii)突变的分子遗传学方法 在病毒RNA中分离出防止组装的物质。无论是在体外还是在 体内组装试验将被用来表征启动 很复杂。病毒核糖核酸序列和构成该病毒的蛋白质(S) 本课程将考察装配的起源。

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.