INTERACTIONS OF MAMMALIAN REOVIRUS PARTICLES W/ EXPRESSED REOVIRUS PROTEINS

该项目的目标是确定结构和 纯化的蛋白质之间形成的蛋白质复合物的功能特性 呼肠孤病毒颗粒和表达的呼肠孤病毒蛋白。 1. 过表达 外衣壳蛋白穆尔和σ 3以一种方式结合呼肠孤病毒核心 在体内重演外衣壳的组装。 的后果 这些结构和构象上的结合相互作用 将确定呼肠孤病毒颗粒的性质。 第一、 将使用负染EM评估样本质量，然后 cryo-SEM以获得高分辨率结构信息。 2. 过表达的呼肠孤病毒外衣壳蛋白sigma 3结合呼肠孤病毒 使这些重组的粒子 功能上类似于病毒体。 重组后的结构 将通过负染EM和低温SEM研究颗粒。 3. 过表达的呼肠孤病毒非结构蛋白muNS结合呼肠孤病毒 丹 这种结合相互作用的影响将由以下决定： 负染电镜和冷冻扫描电镜。 研究这三种病毒蛋白 通过IMR提供的EM技术的组合， 应该产生重要的见解，组装，拆卸， 这种肠道病毒的致病模式，其亲属包括 哺乳动物轮状病毒是鸟类世界婴儿的主要杀手 国家

The goal of this project is to determine the structural and functional properties of protein complexes formed between purified reovirus particles and expressed reovirus proteins. 1. Overexpressed outer-capsid proteins mul and sigma3 bind reovirus cores in a manner that recapitulates outer-capsid assembly in vivo. The consequences of these binding interactions on the structural and conformational properties of the reovirus particle will be ascertained. First, negative-stain EM will be used to assess sample quality, followed by cryo-SEM to obtain high-resolution structural information. 2. Overexpressed reovirus outer-capsid protein sigma3 binds reovirus ISVPs in a fashion that renders these reassembled particles functionally similar to virions. The structurd~of the reassembled particles will be studied by negative-stain EM and cryo-SEM. 3. Overexpressed reovirus nonstructural protein muNS binds reovirus cores. The effects of this binding interaction will be determined by negative-stain EM and cryo-SEM. Studying these three virus-protein complexes by a combination of EM techniques available through the IMR should yield important insights into the assembly, disassembly, and modes of pathogenesis of this enteric virus, whose relatives include the mammalian rotavirus, a major killer of infants in Tbird World countries.