DISSECTING PRION DISEASE MECHANISMS WITH TRANSGENIC MICE

用转基因小鼠剖析朊病毒疾病机制

• 批准号: 2671148
• 负责人: Surachai Supattapone
• 金额: $ 9.29万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-20 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2671148
• 关键词: DNA replication; chemical kinetics; chemical substitution; chimeric proteins; conformation; density gradient ultracentrifugation; endoplasmic reticulum; gene deletion mutation; genetic strain; genetically modified animals; laboratory mouse; mutant; neoplastic cell; nervous system infection; neuroblastoma; pathologic process; prions; protein binding; protein folding; protein isoforms; protein structure function; solubility; species difference; stoichiometry; western blottings

Prion diseases occur in infectious, hereditary, and sporadic forms. Altered conformation of the prion protein (PrP) plays a central role in the pathogenesis of these disorders. My objective is to understand how altered conformations of PrP arise and produce disease. My strategy is to express mutant PrP genes in scrapie-infected neuroblastoma cells and mice, and to study the changes induced by these mutations with biochemical, pathological, and immunological techniques. Two interesting disease models have been generated in preliminary deletional mutagenesis studies: a small infectious prion which can be solubilized in its native state (PrPSc106), and a hereditary neuronal storage disease. In this proposal, I describe how I plan to investigate these disease models further. I will exploit the solubility of PrPSc106 to study the mechanism of prion replication directly. Starting with PrP106 as a minimal backbone, I will investigate the importance of specific PrP epitopes for prion replication, species specificity, and strain properties. Finally, I will determine whether mutant PrP neuronal storage diseases are triggered by protein misfolding or by aberrant retention in the endoplasmic reticulum.

病毒疾病以传染性，遗传和零星形式出现。 prion蛋白（PRP）的改变构象在 这些疾病的发病机理。 我的目标是了解如何 PRP的构象改变并产生疾病。 我的策略是 在crapie感染的神经母细胞瘤细胞中表达突变PRP基因和 小鼠，研究这些突变引起的变化 生化，病理和免疫学技术。 二 有趣的疾病模型是在初步缺失中产生的 诱变研究：一种可以溶解的小型感染性prion 在其本地状态（PRPSC106）和遗传神经元存储 疾病。 在此提案中，我描述了我计划如何调查这些 疾病模型进一步。 我将利用PRPSC106的溶解度 直接研究Prion复制的机制。 从PRP106开始 作为最小的骨干，我将研究特定PRP的重要性 prion复制，物种特异性和应变的表位 特性。最后，我将确定突变的PRP神经元是否 储存疾病是由蛋白质错误折叠或异常引起的 在内质网中保留。