ROLE OF PRION PROTEIN BIOGENESIS IN PATHOGENESIS OF SCRAPIE

朊病毒蛋白生物发生在瘙痒病发病机制中的作用

• 批准号: 6267177
• 负责人: VISHWANATH R LINGAPPA
• 金额: $ 22.24万
• 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: 1998-02-01 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6267177
• 关键词: Xenopus oocyte; cell communication molecule; cell free system; endoplasmic reticulum; genetic translation; genetically modified animals; intracellular transport; laboratory mouse; molecular chaperones; molecular pathology; posttranslational modifications; prions; protein biosynthesis; protein folding; protein reconstitution; protein sequence; protein transport; scrapie; tissue /cell culture

In recent years, progress has been made in dissecting the molecular events of prion protein (PrP) biogenesis. A novel topogenic sequence, termed the Stop Transfer Effector (STE), which directs nascent PrP in cell-free systems to either a doubly transmembrane or a secretory topology, has been identified. The choice between these topologic fates was shown to depend on the presence of a cytosolic factor. An intermediate with features of both topologic forms has been identified in vivo. Pathways by which alternate topologic fates and rapid ER degradation may occur have been identified. However, the relationship of these events to scrapie remains unknown. We propose to explore the role of this novel topogenic sequence in scrapie pathogenesis. The STE sequence will be mutagenized and the effects of mutations on PrP biogenesis and scrapie pathogenesis investigated. Mutants which alter steps in PrP biogenesis will be selected by cell-free transcription- linked translation and Xenopus oocyte microinjection. Some of these will be used to identify receptors and molecular chaperones with which nascent PrP interacts. Selected mutants will be studied in transgenic mice in order to determine if a relationship exists between unusual events in prion biogenesis and the pathogenesis of scrapie. Finally mutants transfected into N2a cells will be used to probe the effect of molecular chaperones on PrP biogenesis and parameters of scrapie infection.

近年来，在分子解剖方面取得了进展。 蛋白质(PrP)的生物发生事件。一种新的拓扑发生序列， 称为停止转移效应器(STE)，它引导新生PrP进入 无细胞系统到双重跨膜或分泌物 拓扑图，已经确定。在这些拓扑命运之间的选择 被证明依赖于胞浆因子的存在。一个 已确定具有这两种拓扑形式特征的中间体 在活体内。拓扑命运交替和快速内质网的途径 已经确定了可能发生的退化。然而，这种关系 这些事件中的哪些对瘙痒来说仍然是未知的。我们建议探讨 这一新的拓扑发生序列在瘙痒病发病机制中的作用。STE 序列将被诱变以及突变对PrP的影响 对瘙痒病的生物发生和发病机制进行了研究。会改变的突变体 PrP生物发生的步骤将通过无细胞转录- 连锁翻译和非洲爪哇卵母细胞显微注射。其中一些人会 用于鉴定新生的受体和分子伴侣 PRP相互作用。选定的突变体将在#年的转基因小鼠中进行研究 中的异常事件之间是否存在关系 朊病毒的生物发生和瘙痒病的发病机制。最后是变种人 将其导入N2a细胞将用于探索分子效应 PrP生物发生中的伴侣蛋白和瘙痒病感染参数。