PROTEIN PROTEIN INTERACTIONS DURING PRION PROTEIN BIOGENESIS

朊病毒蛋白生物发生过程中的蛋白质相互作用

• 批准号: 6299213
• 负责人: VISHWANATH R LINGAPPA
• 金额: $ 22.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6299213
• 关键词: cell free system; cell line; crosslink; disease /disorder model; endoplasmic reticulum; genetically modified animals; immunoprecipitation; intracellular transport; laboratory mouse; molecular pathology; mutant; neural degeneration; phenotype; prions; protein biosynthesis; protein engineering; protein localization; protein protein interaction; protein sequence; protein transport; scrapie; transfection

At the endoplasmic reticulum (ER) membrane, nascent prion protein (PrP) can be synthesized in both fully translocated and transmembrane forms, as a result of the action of Translocation Accessory Factors (TrAFs). The relevance of these observations has recently become apparent with the demonstration of spontaneous neurodegenerative disease in both mice and humans expressing PrP mutations that favor synthesis and export of transmembrane PrP from the ER. Some evidence suggests that the expression of neurodegeneration -associated transmembrane PrP may be a default outcome in the absence of TrAF activity. The investigation of topological regulation during PrP biogenesis is crucial for an understanding the molecular pathogenesis of these genetic prion diseases. Here we propose to dissect the mechanisms by which PrP biogenesis affects PrP topology disease. Here we propose to dissect the mechanism by which PrP biogenesis affects PrP topology, resulting in an transmembrane form that leads to neurodegeneration. Mutations in PrP structural domains will be systematically correlated to effects on topological regulation. Trafficking of selected mutants will be studied in transfected cells. Transgenic mouse model systems will be developed for those mutants with the most interesting phenotypes. Protein-protein interactions between the translocation machinery and nascent PrP will be identified and correlated with the pathway of neurodegeneration. Through this work, the pathway of PrP biogenesis, as it relates to a newly recognized pathway of neurodegeneration, will be better understood in molecular terms. In the long run these studies should make possible novel approaches to manipulation of PrP topology and may contribute to prevention or treatment of prion diseases and other neurodegenerative disease.

在内质网（ER）膜上，由于易位辅助因子（TrAFs）的作用，新生朊蛋白（PrP）可以以完全易位和跨膜形式合成。这些观察结果的相关性最近变得明显，表现出自发性神经退行性疾病的小鼠和人类表达的PrP突变，有利于合成和出口的跨膜PrP从ER。一些证据表明，在缺乏TrAF活性的情况下，神经变性相关的跨膜PrP的表达可能是默认结果。研究PrP生物发生过程中的拓扑调控对于了解这些遗传性朊病毒疾病的分子发病机制至关重要。在这里，我们建议解剖的机制，PrP生物发生影响PrP拓扑结构疾病。在这里，我们建议解剖的机制，PrP生物发生影响PrP拓扑结构，导致跨膜形式，导致神经退行性变。PrP结构域中的突变将系统地与拓扑调控的影响相关。将在转染细胞中研究所选突变体的运输。转基因小鼠模型系统将被开发用于具有最感兴趣的表型的突变体。易位机制和新生PrP之间的蛋白质-蛋白质相互作用将被确定，并与神经变性的途径相关。通过这项工作，PrP生物合成的途径，因为它涉及到一个新认识的神经退行性疾病的途径，将在分子方面得到更好的理解。从长远来看，这些研究应该使新的方法来操纵PrP拓扑结构，并可能有助于预防或治疗朊病毒疾病和其他神经退行性疾病。