Molecular determinants of nodaviral assembly

描述(由申请人提供)：本提案概述了为更好地了解二十面体RNA病毒组装而进行的实验。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的识别、浓缩和封装的方面。它解决了如何识别多部分RNA基因组并将其包装成单个病毒粒子这一悬而未决的问题，并试图确定细胞参数在这一过程中的作用。该实验计划利用了以含有诺达病毒的二十面体RNA为代表的高度发达的模型系统。诺达病毒是一类在遗传、生物化学和结构上都具有异常良好特征的病毒，已被证明对病毒学中的基本问题，包括基因组的颗粒组装和包装的研究非常容易处理。这个 本项目的具体目标是：(1)研究亚细胞区隔在脑损伤中的作用 使用生化和遗传方法的鸡舍病毒(FHV)生命周期；(2)可视化 目的：(1)通过荧光显微镜和电子显微镜研究FHV感染过程中的细胞区划和空间动力学；(3)确定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折叠和凝聚中的作用。

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.