YEAST ESS1, A CONSERVED PPIASE ESSENTIAL FOR MITOSIS

YEAST ESS1，有丝分裂所必需的保守 PPIASE

• 批准号: 6019230
• 负责人: Steven D. HANES
• 金额: $ 13.34万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6019230
• 关键词: Saccharomyces cerevisiae; affinity chromatography; alleles; cell cycle; cell growth regulation; chimeric proteins; enzyme activity; flow cytometry; fungal genetics; gene expression; gene interaction; gene mutation; genetic regulation; immunofluorescence technique; laboratory rabbit; molecular cloning; peptidylprolyl isomerase; phenotype; polymerase chain reaction; protein structure function; regulatory gene; ubiquitin; western blottings; yeast two hybrid system

Yeast and human cells require the ESS1 gene in order to divide. This essential gene controls cell cycle progression through mitosis, and given its extraordinary conservation, is probably required in all eukaryotes. The goal of this work is to understand how ESS1 controls the cell cycle. The yeast saccharomyces cerevisiae, the organism in which ESS1 was discovered, will be used as a model system. ESS1 is highly conserved both structurally and functionally; ESS1 homologs from Drosophila melanogaster and humans rescue yeast ess1 mutants. The Ess1 protein and its counterparts are the only known proteins to contain both a WW domain and a PPIase (peptidyl-prolyl isomerase) domain. The WW domain is a protein-protein interaction module found in proteins such as Dystrophin, encoded by the human muscular dystrophy gene. The PPIase domain is a catalytic module that controls the folding and activity of kinases, receptors and transcription factors and is found in proteins like cyclophilin that mediate the effects of immunosuppressive drugs. How these domains function in Ess1 to control the cell cycle is not known. In this proposal, yeast genetics will be used understand how Ess1 controls mitotic progression and how the WW and PPIase domains contribute to this activity. This will be accomplished by (1) conducting an extensive structure- function analysis of the Ess1 WW and PPIase domains, (2) by isolating conditional alleles of ESS1 gene and using these alleles for cell cycle analysis, and (3) by identifying other components of the ESS1 pathway using suppressor screens and tow-hybrid selections. An understanding of how ESS1 functions in yeast will illuminate poorly understood pathways of mitotic regulation and will provide insight into the control of the cell cycle in all eukaryotic cells.

酵母和人类细胞需要ESS1基因才能分裂。 该基因通过有丝分裂控制细胞周期的进展， 鉴于其非凡的保护，可能是所有人都需要的 真核生物。 这项工作的目的是了解ESS1 控制细胞周期。 酿酒酵母的酵母菌， 发现ESS1的有机体将用作模型 系统。 ESS1在结构上都是高度保守的， 在功能上；果蝇Melanogaster的ESS1同源物和 人类营救酵母ESS1突变体。 ESS1蛋白及其 对应物是唯一包含WW的唯一已知蛋白质 结构域和PPIASE（肽基 - 蛋白酶异构酶）结构域。 WW 域是在蛋白质中发现的蛋白质 - 蛋白质相互作用模块 例如肌营养不良蛋白，由人类肌肉营养不良编码 基因。 PPIASE结构域是一个控制该模块的催化模块 激酶，受体和转录因子的折叠和活性 并在诸如环磷脂之类的蛋白质中发现，介导 免疫抑制药物。 这些域如何在ESS1中起作用 控制细胞周期尚不清楚。 在此提案中，酵母遗传学 将使用将了解ESS1如何控制有丝分裂进展和 WW和PPIASE领域如何促进此活动。 这 将通过（1）进行广泛的结构来完成 - ESS1 WW和PPIASE域的功能分析，（2） 隔离ESS1基因的条件等位基因，并将这些等位基因用于 细胞周期分析，（3）通过识别其他组件 ESS1使用抑制器屏幕和拖曳杂交的选择。 对ES1在酵母中的功能的理解将阐明 知识较低的有丝分裂调节途径，并将提供 深入了解所有真核细胞中细胞周期的控制。