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: 6138698
• 负责人: YURY O CHERNOFF
• 金额: $ 14.83万
• 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 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6138698
• 关键词: SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; cytoskeletal proteins; extracellular matrix proteins; fluorescence microscopy; fungal proteins; genetic screening; immunoprecipitation; intracellular; molecular assembly /self assembly; molecular chaperones; nucleic acid sequence; polymerase chain reaction; prions; protein isoforms; protein protein interaction; scintillation counter; southern blotting; yeast two hybrid system

The transmissible neurodegenerative encephalopathies (called prion diseases) in mammals and humans, as well as age-related pathologies such as Alzheimer's syndrome, are associated with a nucleated polymerization" process leading to the formation of protein aggregates. Prion diseases and Alzheimer's syndrome are fatal and incurable. The "protein only" model of prion diseases suggests that abnormal prion protein may serve as an infectious agent by "seeding" formation of the new prions. Recent finding of prion-like proteins in the yeast Saccharomyces cerevisiae suggests a wide distribution and possible biological importance of prion phenomena. From the geneticist' s point of view, the "protein only" model of prion phenomena suggests that, in addition to the information which is coded by the sequence, some protein and/or multiprotein structures contain additional information which is "coded" in the structure itself. Highly ordered cellular structures such as cytoskeletal formations, are the most likely candidates for the systems of "structural" coding. The proposed project is designed to test a hypothesis, which states that the appearance of prion aggregates results from an abortive form of the structure-forming process, normally leading to assembly of the ordered intracellular or extracellular multiprotein complexes. The yeast prion- forming translational release factor Sup35p (eRF3) will be used as an experimental model. Roles of the chaperone helpers and cytoskeleton- associated proteins in formation and propagation of the Sup35pPSI prion, and in the cytoskeletal assembly will be compared by using genetic and biochemical approaches. Protein-protein interactions, which involve the prion-forming domain of the Sup35p, will be characterized. Results obtained will shed light on the mechanisms, integrating the potential prion-forming processes into normal cellular physiology, and may eventually lead to the development of new cures for prion diseases and related disorders.

传染性神经退行性脑病（称为朊病毒 疾病），以及与年龄相关的病理学， 如阿尔茨海默氏综合征，与核聚合有关。 导致蛋白质聚集体形成的过程。 朊病毒病 和老年痴呆症都是致命且无法治愈的 “只有蛋白质” 朊病毒疾病的模型表明，异常朊病毒蛋白可能 作为一种传染媒介，通过“播种”形成新的朊病毒。 朊病毒样蛋白在酵母菌中的发现 酿酒酵母表明广泛的分布和可能的生物学 朊病毒现象的重要性。 从遗传学家的观点来看， 朊病毒现象的“仅蛋白质”模型表明，此外， 对于由序列编码的信息，一些蛋白质和/或 多蛋白质结构包含额外的信息， 在结构本身。 高度有序的细胞结构，如 细胞骨架的形成，是最有可能的候选人的系统 “结构性”编码。 拟议的项目旨在检验一个假设，该假设指出， 朊病毒聚集体的出现是由于 结构形成过程，通常导致有序的组装 细胞内或细胞外多蛋白复合物。 酵母朊病毒- 形成翻译释放因子Sup 35 p（eRF 3）将被用作 实验模型 分子伴侣和细胞骨架的作用- Sup 35 pPSI朊病毒形成和繁殖中的相关蛋白， 和细胞骨架组装将通过使用遗传和 生物化学方法。 蛋白质间的相互作用， Sup 35 p的朊病毒形成结构域将被表征。 结果 获得将阐明的机制，整合的潜力 朊病毒形成过程进入正常的细胞生理学，并可能 最终导致朊病毒疾病的新疗法的发展， 相关疾病。