REGULATION OF MALTOSE FERMENTATION IN SACCHAROMYCES

酵母菌中麦芽糖发酵的调控

• 批准号: 2175128
• 负责人: Corinne Anthony Michels
• 金额: $ 28.64万
• 依托单位: QUEENS COLLEGE
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-07-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2175128
• 关键词: Saccharomyces; alpha glucosidase; cell cycle; chimeric proteins; enzyme induction /repression; enzyme structure; eukaryote; fermentation; fungal genetics; galactokinase; gene deletion mutation; gene expression; genetic mapping; genetic strain; glucose; maltose; mutant; nucleic acid hybridization; nucleic acid sequence; phosphorylation; posttranslational modifications; protein biosynthesis; radiotracer; structural genes; sucrose; transcription factor; transport proteins

The long range goal of this project is to identify and characterize molecular mechanisms regulating gene expression in eukaryotes. Maltose fermentation in the Saccharomyces yeasts has been chosen as a model regulated system. The initial steps of maltose fermentation are carried out by maltose permease (the transport protein) and maltase (the cleavage enzyme). Synthesis of these enzymes is induced by maltose and repressed by glucose and both regulatory processes are controlled at the level of transcription. Our studies have identified a trans-acting positive factor, the MAL-activator protein, which mediates the transcriptional regulation of the structural genes encoding maltose permease and maltase. Maltose fermenting Saccharomyces strains contain at least one of a series of complex loci (MAL1, MAL2, MAL3, MAL4 and MAL6) each of which encodes maltose permease, maltase and the MAL-activator. The experiments described in this proposal are designed to reveal the details of the role played by the MAL-activator in regulating expression of the MAL structural genes. For this, we will a) complete our analysis of the functional domains of the MAL-activator using noninducible and constitutive mutations by in vitro and in vivo mutagenesis techniques; b) identify proteins which interact with the MAL-activator and play a role in maltose induction using the "two hybrid protein system" and c) characterize the role of phosphorylation and/or proteolytic degradation of the MAL-activator in the regulation of fermentation. We will investigate the factors involved in regulating the synthesis of the MAL-activator itself by identifying upstream controlling elements of the MAL-activator gene. We will explore the role of MIG1 protein in the glucose repression of the MAL gene, both structural and regulatory. Finally, we plan to reinvestigate the reported position effect controlling the timed synthesis of maltase during the yeast cell cycle.

该项目的远距离目标是识别和表征 调节真核生物基因表达的分子机制。 麦芽糖 酵母酵母中的发酵已被选为模型 受管制系统。 麦芽糖发酵的最初步骤被携带 通过麦芽糖渗透酶（转运蛋白）和麦芽糖酶（裂解 酶）。 这些酶的合成是由麦芽糖诱导的，并由 葡萄糖和两个调节过程均受控制 转录。 我们的研究已经确定了跨性别的积极因素， MAL激活蛋白介导的转录调节 编码麦芽糖渗透酶和麦芽糖酶的结构基因。 麦芽糖 发酵酵母菌株至少包含一系列的一系列 复杂的基因座（Mal1，Mal2，Mal3，Mal4和Mal6）每个编码 麦芽糖渗透酶，麦芽糖酶和麦芽活化剂。 描述的实验 在此提案中旨在揭示作用的细节 在调节MAL结构基因表达方面的MAL激活剂。 为此，我们将a）完成对该功能域的分析 通过体外和不可诱导和组成型突变的MAL激活剂 体内诱变技术； b）确定与之相互作用的蛋白质 Mal-Acrovator并使用“两个”在麦芽糖诱导中发挥作用 杂化蛋白系统”和c）表征磷酸化的作用 和/或蛋白水解降解在调节中 发酵。 我们将调查调节的因素 通过识别上游控制的MAL激活剂本身的合成 MAL激活基因的元素。 我们将探索MIG1的作用 MAL基因的葡萄糖抑制中的蛋白质，包括结构和结构 监管。 最后，我们计划重新投资报告的位置效应 控制酵母细胞周期期间麦芽糖酶的定时合成。