INTERACTIONS OF MAMMALIAN REOVIRUS PARTICLES W/ EXPRESSED REOVIRUS PROTEINS

该项目的目标是确定结构和 纯化后的蛋白质复合体的功能性质 呼肠孤病毒颗粒和表达的呼肠孤病毒蛋白。1.过度表达 外壳蛋白mu1和sigma3以某种方式与呼肠孤病毒核心结合 这概括了体内的外衣壳组装。其后果是 结构和构象上的这些结合作用 呼肠孤病毒颗粒的性质将被确定。第一, 将使用负染色EM来评估样本质量，然后是 低温扫描电子显微镜以获得高分辨率的结构信息。2. 过表达的呼肠孤病毒外壳蛋白Sigma3与呼肠孤病毒结合 ISVP以一种方式呈现这些重新组装的粒子 在功能上类似于病毒粒子。重组的结构~ 用负染色电子显微镜和低温扫描电子显微镜对颗粒进行研究。3. 过表达的呼肠孤病毒非结构蛋白MUNS与呼肠孤病毒结合 核心。这种绑定相互作用的效果将由以下因素决定 负染EM和冷冻扫描电子显微镜。对这三种病毒蛋白的研究 通过IMR提供的EM技术的组合生成复合体 应该对组装、拆卸和 这种肠道病毒的致病模式包括其近亲 哺乳动物轮状病毒，三鸟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.