ANALYSIS OF SPHERICAL VIRUS ASSEMBLY

球形病毒的组装还不清楚。 客观 这一建议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）。 的 这项建议的具体目标是：（一）界定和绘制病毒 负责启动病毒装配的RNA和外壳蛋白序列; (ii)表征起始复合物的生物化学性质， 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是必不可少的，以及（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。 远景目标 这项研究的目的是确定 病毒组装 对病毒体组装的理解可能揭示 干扰组装，提供了新的方法来控制 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， 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。 装配起始复合物 将以三种互补方法为特征：（i）生物化学 一种试图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 在病毒RNA被分离，阻止组装。 在体外和 将使用体内组装测定来表征引发 复杂. 病毒RNA序列和构成病毒RNA序列的蛋白质两者都可以被识别。 将检查装配的起源。

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.