Molecular Analysis of Yeast Prions

自从Reed Wickner首次提出酿酒酵母中鲜为人知的非孟德尔状态以来，[URE3] 和[PST]，是由内源蛋白、真菌和真菌中的[PST]蛋白类病毒转化而成的 特别是，已被证明是探索普赖恩生物学普遍特征的独特而强大的系统。一个 很少有主要的见解包括直接证明普里恩遗传的纯蛋白质假说， 侵染性构象中被编码的PrP菌株的分子基础的阐明 蛋白质，发现一种新的Pron蛋白的增殖，并暗示基于Pron的遗传 可能是一种普遍存在的蛋白质功能表观遗传控制机制，并揭示了宿主 伴侣机制在催化病毒复制中起着至关重要的作用。与所有实验一样，这些 研究结果还提出了一系列基本问题，如不同菌株的结构基础 构象，这些不同的构象是如何遗传的，为什么它们对细胞的 生理学，也许最深刻的是，普恩的生物学作用是什么。我们现在有了工具和 知识基金会开始为这些问题提供明确和具体的答案。其中的这些见解 TURN应该直接为理解哺乳动物PrP的PrP传染性原理的努力提供信息 更广泛地说，为了促进我们对蛋白质如何错误折叠以及为什么错误折叠以及如何错误折叠的理解 错误折叠的形式会影响处于疾病和非疾病状态的细胞。 为了实现这一目标，我们建议重点关注以下三个方面： 定义Pron变种的结构基础。 定义Prion的一级结构如何决定首选变种的光谱 使用核糖体图谱来确定[PST]Pron的生理影响以及Pron菌株如何 酵母的构象和遗传背景调节了这些效应。 相关性(请参阅说明)： 蛋白质错误折叠是一系列神经退行性疾病的标志，包括Pron疾病和 更常见的非传染性疾病，如阿尔茨海默病和帕金森。

Since Reed Wickner first proposed that otherwise obscure non-Mendelian states in S. cerevisiae, [URE3] and [PST], result from prion-like conversions of endogenous proteins, fungal prions in general and [PST] in particular, have proven to be uniquely powerful systems for exploring universal features of prion biology. A few ofthe major insights include direct demonstrafion ofthe protein only hypothesis of prion inheritance, elucidafion of molecular basis of prion strains as being enciphered in the conformation ofthe infectious protein, discovery of a proliferation of novel prion proteins and the suggestion that prion-based inheritance could be a ubiquitous mechanism for epigenetic control of protein function, and revelation that the host chaperone machinery plays a crifical role in catalyzing prion replication. As with all experiments, these findings also raise a host of fundamental questions such as the structural basis ofthe different strain conformations, how these different conformations are inherited and why do they differentially impact a cell's physiology, and perhaps most profoundly what is the biological role of prions. We now have the tools and the intellectual foundafion to begin to provide clear and concrete answers to these questions. These insights in turn should directly inform efforts to understand the principles of prion infectivity for the mammalian PrP protein and more generally to advance our understanding of how and why proteins misfold and how such misfolded forms impact a cell in both disease and non disease states. To accomplish this, we propose to focus on the following three areas: Define the structural basis of prion strain variants. Define how the primary structure of prion determines the spectrum of preferred strain variants Use ribosome profiling to define the physiological impact ofthe [PST] prion and how the prion strain conformation as well as the genetic background ofthe yeast modulate these effects. RELEVANCE (See instructions): Protein misfolding is a hallmark of a wide range of neurodegenerative disorders including prion diseases and far more common noninfectious disease such as Alzheimers and Parkinson.