GLUCOSE INACTIVATION OF MALTOSE PERMEASE IN YEAST

酵母中麦芽糖渗透酶的葡萄糖失活

• 批准号: 2186863
• 负责人: Corinne Anthony Michels
• 金额: $ 16.1万
• 依托单位: QUEENS COLLEGE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2186863
• 关键词: Saccharomyces; biological signal transduction; carbohydrate transport; cyclic AMP; enzyme activity; enzyme induction /repression; fermentation; fluorescence microscopy; fungal genetics; gene expression; glucose; high performance liquid chromatography; immunoprecipitation; maltose; membrane transport proteins; microorganism metabolism; mutant; nucleic acid sequence; permease; phosphorylation; posttranslational modifications; proteolysis; radiotracer; transcription factor; ubiquitin; western blottings

Maltose fermentation in Saccharomyces is initiated by its active transport into the cell by maltose permease. The level of activity of maltose permease is under dual regulation. Firstly, transcription of the gene encoding maltose permease is induced by maltose and repressed by glucose. Secondly, maltose permease is glucose inactivated, that is, the ability to transport maltose is rapidly lost following the addition of glucose to a fermenting culture. We propose to determine the mechanism of this post-translational regulation of maltose permease. MA161 encodes maltose permease in Saccharomyces. We have inserted an epitope-tag at the N-terminal end of the Ma161 protein to enable us to follow the fate of the protein during glucose-induced inactivation by using techniques such as immunoprecipitation, immunofluorescence microscopy and Western analysis. The projects described in this proposal explore the possibility that maltose permease is modified by phosphorylation and/or proteolytically degraded during inactivation. We will use the tools of Saccharomyces genetics and the expanding number of mutant strains with alterations affecting the secretory, signal transduction and glucose-response pathways to determine the gene products utilized for the inactivation process. We will isolate glucose inactivation resistant mutations in MA161 and define new Saccharomyces genes encoding products involved in glucose-induced inactivation.

葡萄糖中的麦芽糖发酵是由其活性开始的 通过麦芽糖粘酶运输到细胞中。 活动水平 麦芽糖渗透率受双重调节。 首先，转录 编码麦芽糖渗透酶的基因是由麦芽糖诱导的，并被抑制 葡萄糖。 其次，麦芽糖渗透酶被葡萄糖灭活，即 添加后，运输麦芽糖的能力迅速失去了 葡萄糖到发酵培养物。 我们建议确定机制 麦芽糖渗透酶的翻译后调节。 MA161编码糖疗中的麦芽糖渗透酶。 我们插入了 MA161蛋白的N末端的表位标签使我们能够 在葡萄糖诱导的失活过程中遵循蛋白质的命运 使用免疫沉淀，免疫荧光等技术 显微镜和西方分析。 本提案中描述的项目 探索Maltose Perease修改的可能性 在失活期间磷酸化和/或蛋白水解降解。 我们 将使用糖类遗传学的工具和扩大的数量 突变菌株具有影响分泌，信号的变化 转导和葡萄糖反应途径以确定基因产物 用于灭活过程。 我们将分离葡萄糖 MA161中的抗活化突变并定义新的糖疗法 编码参与葡萄糖诱导的失活的基因。