PROTEIN PROTEIN INTERACTIONS DURING PRION PROTEIN BIOGENESIS

在内质网(ER)膜上，由于易位辅助因子(TRAF)的作用，新生蛋白(PrP)可以完全易位和跨膜两种形式合成。这些观察结果的相关性最近变得明显，因为在小鼠和人类中都有自发的神经退行性疾病表达PrP突变，这些突变有利于从内质网合成和输出PrP。一些证据表明，在缺乏Traf活性的情况下，神经退行性变相关的跨膜PrP的表达可能是一种默认结果。研究PrP生物发生过程中的拓扑调控对于理解这些遗传性PrP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、导致跨膜形式导致神经退化的机制。PrP结构域的突变将与拓扑调控的影响系统相关。将在转基因细胞中研究所选突变体的转运。将为那些具有最有趣表型的突变体开发转基因小鼠模型系统。易位机制和新生PrP之间的蛋白质-蛋白质相互作用将被识别，并与神经退化的途径相关。通过这项工作，PrP的生物发生途径将从分子角度得到更好的理解，因为它与一种新发现的神经退变途径有关。从长远来看，这些研究将使操纵PrP拓扑学的新方法成为可能，并可能有助于预防或治疗PrP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.