PROTEIN PROTEIN INTERACTIONS DURING PRION PROTEIN BIOGENESIS

在内质网（ER）膜上，由于易位辅助因子（TrAFs）的作用，新生朊病毒蛋白（PrP）可以以完全易位和跨膜的形式合成。随着自发性神经退行性疾病在小鼠和人类中表达有利于内质网跨膜PrP合成和输出的PrP突变，这些观察结果的相关性最近变得明显。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的表达可能是默认的结果。研究PrP生物发生过程中的拓扑调控对于理解这些遗传朊病毒疾病的分子发病机制至关重要。在这里，我们建议剖析PrP生物发生影响PrP拓扑疾病的机制。在这里，我们建议剖析PrP生物发生影响PrP拓扑结构的机制，从而导致导致神经变性的跨膜形式。PrP结构域的突变将系统地与拓扑调节的影响相关。将在转染的细胞中研究选定突变体的贩运。转基因小鼠模型系统将用于那些具有最有趣表型的突变体。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，并与神经退行性变的途径相关。通过这项工作，PrP的生物发生途径，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，这些研究应该可以为PrP拓扑的操作提供新的方法，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。

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.