Genetic Study of the Yeast Prion-Interacting Proteins

酵母朊病毒相互作用蛋白的遗传学研究

• 批准号: 6798178
• 负责人: YURY O CHERNOFF
• 金额: $ 26.54万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6798178
• 关键词: SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; affinity chromatography; cytoskeletal proteins; extracellular matrix proteins; fluorescence microscopy; fungal proteins; heat shock proteins; immunoprecipitation; molecular assembly /self assembly; molecular chaperones; molecular cloning; nucleic acid sequence; polymerase chain reaction; prions; protein isoforms; protein protein interaction; southern blotting; ubiquitin; yeast two hybrid system

DESCRIPTION (provided by applicant): Incurable infectious neurodegenerative diseases, such as "mad cow" disease and human Creutzefeldt-Jacob disease, are postulated to be transmitted by an abnormal aggregation-prone protein isoform (prion). Prion aggregates are believed to be capable of seeding aggregation of the normal cellular protein, converting it into a prion. Pathology of prion diseases is reminiscent of other protein assembly disorders (such as Alzheimer?s disease) associated with amyloid-like aggregation. In yeast, prions control phenotypic traits inherited via cytoplasm. This provides a molecular basis for protein-based inheritance. As several unrelated proteins exhibit prion-forming potential, it is likely that protein-based inheritance may play an important biological role. The power of yeast genetic analysis makes yeast a useful model for studying general mechanisms of prion propagation. While many proteins can form amyloid-like aggregates in vitro, only some of them are capable of transmitting the aggregated state in vivo. This means that cells are normally able to prevent aggregate propagation. However, prions are overcoming the cellular defense systems. Moreover, our research has proven that the cellular stress-defense machinery becomes an essential component of prion propagation in the yeast cell. The overall, goal of this proposal is to uncover mechanisms by which cellular systems control prion formation and propagation in the yeast cell. This will explain how environmental and physiological factors induce protein-based inheritable variations, and may provide a new tool for curing aggregation-based disorders by altering the cellular regulatory systems. Proteins affecting prion formation and propagation in yeast will be investigated: stress-related chaperones of the evolutionary conserved Hsp100 and Hsp70 groups; ubiquitin system, normally involved in targeting misfolded proteins for degradation; and cytoskeletal assembly proteins involved in formation of cortical actin patches and endocytic vesicles. Specific Aims of the proposal are as follows: 1) To study a mechanism of prion regulation by the heat shock protein balance. 2) To study the role of ubiquitin system in the cellular control of yeast prions. 3) To study interactions between prions and cytoskeleton-associated structures in yeast.

描述（由申请人提供）：无法治愈的感染性神经退行性 疾病，例如“疯牛”疾病和人类creutzefeldt-jacob疾病，是 假设将通过异常骨聚集的蛋白质同工型传播 （prion）。据信，prion骨聚集体能够播种 正常细胞蛋白，将其转化为prion。病毒的病理学 疾病让人想起其他蛋白质组装疾病（例如 阿尔茨海默氏病）与淀粉样蛋白样聚集有关。在酵母中，prions 通过细胞质遗传的控制表型特征。这提供了分子 基于蛋白质的遗传基础。由于几种无关蛋白质展示 基于蛋白质的遗传可能会发挥作用 重要的生物学作用。酵母遗传分析的力量使酵母成为 用于研究普通传播的一般机制的有用模型。而很多 蛋白质可以在体外形成淀粉样蛋白样骨料，其中一些是 能够在体内传输汇总状态。这意味着细胞是 通常能够防止总体传播。但是，王室正在克服 蜂窝防御系统。此外，我们的研究证明了 细胞应力防御机械成为prion的重要组成部分 酵母细胞中的传播。该提案的总体目标是揭示蜂窝系统控制Prion的形成和传播的机制 酵母细胞。这将解释环境和生理因素如何 诱导基于蛋白质的遗传变体，并可能为 通过改变细胞调节系统来治愈基于聚集的疾病。 影响酵母菌prion形成和传播的蛋白质将是 研究：进化保守的与应力相关的伴侣 HSP100和HSP70组；泛素系统，通常参与靶向 错误折叠的蛋白质降解；和细胞骨架组件蛋白涉及 在皮质肌动蛋白斑块和内吞囊泡的形成中。具体目标 该提议如下：1）研究通过 热休克蛋白平衡。 2）研究泛素系统在 酵母菌王室的细胞控制。 3）研究私人和 酵母中的细胞骨架相关结构。