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.

朊病毒病以传染性、遗传性和散发性形式发生。 朊病毒蛋白 (PrP) 构象的改变在 这些疾病的发病机制。 我的目标是了解如何 PrP 构象发生改变并产生疾病。 我的策略是 在痒病感染的神经母细胞瘤细胞中表达突变的 PrP 基因 小鼠，并研究这些突变引起的变化 生物化学、病理学和免疫学技术。 二 有趣的疾病模型已经在初步删除中产生 诱变研究：一种可溶解的小型传染性朊病毒 其天然状态（PrPSc106）和遗传性神经元存储 疾病。 在这个提案中，我描述了我计划如何调查这些 进一步建立疾病模型。 我将利用 PrPSc106 的溶解度来 直接研究朊病毒复制机制。 从 PrP106 开始 作为最低限度的骨干，我将研究特定 PrP 的重要性 朊病毒复制、物种特异性和菌株的表位 特性。最后，我将确定是否存在突变的 PrP 神经元 储存疾病是由蛋白质错误折叠或异常引起的 滞留在内质网中。