Molecular determinants of nodaviral assembly

描述（由申请人提供）：本提案概述了旨在更好地理解二十面体RNA病毒组装的实验。 它侧重于衣壳蛋白和病毒RNA之间的特定相互作用，以及控制病毒基因组识别，浓缩和纯化的方面。 它解决了尚未解决的问题，如何识别和包装成一个单一的病毒体的多组分RNA基因组，它试图定义在这个过程中的细胞参数的作用。该实验计划利用了以含有二十面体RNA的野田病毒为代表的高度发达的模型系统。 野田病毒是一个在遗传、生物化学和结构上非常好表征的病毒家族，已证明其对于研究病毒学中的基本问题（包括基因组的颗粒组装和包装）是高度易处理的。 的 本项目的具体目标是：（1）研究亚细胞区室化在细胞凋亡中的作用。 利用生物化学和遗传学方法研究鸡群病毒（FHV）的生命周期;（2）可视化 通过荧光显微镜和电子显微镜观察FHV感染过程中的细胞区室化和空间动力学;（3）鉴定FHV基因组包装中涉及的机制策略和分子元件;（4）确定FHV外壳蛋白在组装过程中病毒RNA折叠和缩合中的作用。

DESCRIPTION (provided by applicant): This proposal outlines experiments to develop a better understanding of icosahedral RNA virus assembly. It focuses on specific interactions between capsid protein and viral RNA as well as aspects that control recognition, condensation and encapsidation of the viral genome. It addresses the unresolved issue of how multipartite RNA genomes are recognized and packaged into a single virion and it seeks to define the role of cellular parameters in this process. The experimental plan takes advantage of a highly developed model system represented by the icosahedral RNA containing nodaviruses. Nodaviruses are a family of genetically, biochemically and structurally exceptionally well characterized viruses that have proven to be highly tractable for the study of fundamental questions in virology, including particle assembly and packaging of the genome. The specific aims of this project are: (1) to investigate the role of subcellular compartmentalization in the Flock House Virus (FHV) life cycle using biochemical and genetic approaches; (2) to visualize cellular compartmentalization and spatial dynamics during FHV infection by fluorescence microscopy and electron microscopy; (3) to identify mechanistic strategies and molecular elements involved in packaging of the FHV genome and (4) to define the role of FHV coat protein in folding and condensation of viral RNA during assembly.