ANALYSIS OF CELL CYCLE UBIQUITIN CONJUGATION IN YEAST

酵母细胞周期泛素结合分析

• 批准号: 3468341
• 负责人: MARK G. GOEBL
• 金额: $ 9.8万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 1995-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3468341
• 关键词: Saccharomyces cerevisiae; alleles; aminoacid; cell cycle; cell growth regulation; cytogenetics; drug metabolism; enzyme complex; enzyme induction /repression; enzyme mechanism; enzyme substrate; fungal genetics; growth factor receptors; in situ hybridization; mutant; neural degeneration; nucleic acid sequence; posttranslational modifications; protein biosynthesis; ubiquitin; yeasts

The long range goal of the proposed research is to understand the role of ubiquitin conjugation in the regulation of early cell cycle events. This project will have two primary objectives. The first objective will be a biochemical and genetic characterization of the CDC34 ubiquitin-conjugating enzyme of yeast. The second objective will be to identify new proteins that act as regulators or substrates of the CDC34 ubiquitin-conjugating enzyme. The well developed genetics of yeast make it an ideal system in which to determine the importance of biochemically defined protein domains and modification events through mutational analysis. The yeast system is also amenable to the identification of interacting proteins through the isolation of mutations and their subsequent molecular analysis. First, a variety of biochemical techniques will be used to verify the identity of several putative isoforms of the CDC34 protein and to then characterize the structural differences between the isoforms and to use in vitro mutagenesis to determine the importance of any structural differences. Secondly, the genetic interaction known as synthetic lethality will be used as the basis of a screen to isolate mutations in genes encoding proteins that interact as regulators or substrates of the CDC34 enzyme. The mutants will be examined for affects on the cell division cycle and the function of the CDC34 enzyme. A better understanding of ubiquitin metabolism may provide insights into a number of diseases and health-related questions. Ubiquitin is found in the neuronal inclusions associated with several degenerative neurological diseases. It is also found attached to a number of growth factor receptors. Thus an enhanced understanding of ubiquitin function may improve our knowledge in areas ranging from cell growth control to neurodegenerative diseases.

拟议研究的远距离目标是了解 早期细胞周期事件调节中的泛素结合。 这 项目将有两个主要目标。 第一个目标将是 Cdc34泛素结合的生化和遗传表征 酵母的酶。 第二个目标是识别新蛋白质 它充当Cdc34泛素结合的调节剂或底物 酶。 发达的酵母遗传学使其成为理想的系统 确定生化定义的蛋白质结构域的重要性 和通过突变分析进行修改事件。 酵母系统是 也可以通过鉴定通过 突变的分离及其随后的分子分析。 首先，将使用多种生化技术来验证 Cdc34蛋白的几种推定同工型的身份，然后 表征同工型之间的结构差异并在 体外诱变以确定任何结构的重要性 差异。 其次，称为合成的遗传相互作用 致死性将被用作隔离突变的屏幕基础 编码作为调节剂或底物相互作用的蛋白质的基因 Cdc34酶。 将检查突变体对细胞的影响 Cdc34酶的分裂周期和功能。 更好地了解泛素代谢可能会提供对 疾病和与健康有关的问题的数量。 泛素在 与几个退化性神经学有关的神经元内包含物 疾病。 还发现它附在许多生长因子上 受体。 因此，增强对泛素功能的理解可能 提高我们在细胞生长控制到的领域知识 神经退行性疾病。