MECHANISMS BY WHICH ROTAVIRUS INDUCES DIARRHEA

这项奖学金申请提出了研究，以了解致病性 轮状病毒引起儿童严重腹泻的机制。 轮状病毒具有独特的形态发生，其中颗粒获得 由新形成的细胞出芽形成的短暂的膜包膜。 亚病毒颗粒进入内质网（ER）。这个过程是 由称为NSP 4的病毒非结构糖蛋白介导， 定位于受感染细胞的内质网膜。最近的研究表明， 实验室已经显示NSP 4和合成肽NSP 4 114.135 （对应于NSP 4氨基酸114-135），当注射时 腹腔内或腹腔内，诱导幼鼠腹泻， 大鼠NSP 4和NSP 4 114.1350，当表达于或外源添加至 昆虫细胞，调动细胞内钙。我们推测NSP 4可能是 引起腹泻的毒力因子 NSP 4的钙动员特性可能 是一种信号转导途径的基础， 致病性电生理学方法以及传统的 在本申请中提出了分子生物学技术， 描述NSP 4动员细胞内钙的机制， 引起幼畜腹泻。了解这些机制可能 为解决轮状病毒的致病途径提供了关键。因为 这种轮状病毒非结构糖蛋白的独特性质，我们 有可能阐明病毒致病的新机制， 病毒引起钙动员的信号转导途径 proteins.在拟议的研究中，我将学习新技术，包括： Ussing小室，平面脂双层，重组体的构建和应用 质粒或病毒、点特异性突变和其他分子方法 必须进行研究，以回答基本问题， 分子生物学

This fellowship application proposes studies to understand the pathogenic mechanism by which rotavirus causes severe diarrhea in children. Rotavirnses have a unique morphogenesis in which particles obtain a transient membrane envelope that is formed by the budding of newly made subviral particles into the endoplasmic reticulum (ER). This process is mediated by a viral nonstructural glycoprotein called NSP4, which is localized to the ER membrane of infected cells. Recent studies in this laboratory have shown that NSP4 and a synthetic peptide NSP4 114.135 (corresponding to NSP4 amino acids 114-135), when injected intraperitoneally or intraduodenally, induced diarrhea in young mice and rats. NSP4 and NSP4 114.1350, when expressed in or added exogenously to insect cells, mobilize intracellular calcium. We speculate that NSP4 may be the virulence factor responsible for inducing the diarrhea caused by rotaviruses, and that the calcium mobilization property of NSP4 may underlie a signal transduction pathway which contributes to virus pathogenicity. Electrophysiological approaches as well as conventional molecular biological techniques are proposed in this application to delineate the mechanisms by which NSP4 mobilizes intracellular calcium and induces diarrhea in young animals. Understanding these mechanisms may provide the key to resolving the pathogenic pathway of rotaviruses. Because of the unique properties of this rotavirus nonstructural glycoprotein, we are likely to elucidate new mechanisms of viral pathogenesis and novel signal transduction pathways of calcium mobilization caused by viral proteins. In the proposed research, I will learn new techniques including: Ussing chamber, planar lipid bilayer, construction and use of recombinant plasmids or virus, point specific mutations, and other molecular approaches necessary to carry out research to answer fundamental questions in molecular biology.