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.

可传播的神经退行性脑病（称为prion） 哺乳动物和人类的疾病）以及与年龄有关的病理 正如阿尔茨海默氏症综合征一样，与成核聚合有关” 过程导致蛋白质聚集体的形成。 病毒疾病 阿尔茨海默氏症综合征是致命的，无法治愈的。 “仅蛋白质” 王室疾病的模型表明异常的prion蛋白可以使用 通过“播种”新奖励的形成作为感染力。 最近发现酵母酵母中类似蛋白质的蛋白 酿酒酵母提出了广泛的分布和可能的生物学 王室现象的重要性。 从遗传学家的角度来看， prion现象的“仅蛋白质”模型表明，此外 根据序列编码的信息，某些蛋白质和/或 多蛋白结构包含“编码”的其他信息 在结构本身中。 高度有序的细胞结构，例如 细胞骨架形成是系统最有可能的候选者 “结构”编码。 拟议的项目旨在检验假设，该假设指出 prion骨料的出现是由堕胎形式的 结构形成过程，通常导致秩序的组装 细胞内或细胞外多蛋白复合物。 酵母菌 - 形成翻译释放因子SUP35P（ERF3）将用作 实验模型。 伴侣助手和细胞骨架的角色 相关的蛋白质在sup35ppsi prion的形成和传播中， 在细胞骨架中，将通过使用遗传和 生化方法。 蛋白质 - 蛋白质相互作用，涉及 SUP35P的prion形成域将被表征。 结果 获得的将阐明机制，使电势整合 培养在正常细胞生理学的过程中，可能 最终导致开发新的病毒疾病治疗方法 相关疾病。