REGULATION OF MALTOSE FERMENTATION IN SACCHAROMYCES

酵母菌中麦芽糖发酵的调控

• 批准号: 2900531
• 负责人: Corinne Anthony Michels
• 金额: $ 31.52万
• 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 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2900531
• 关键词: Saccharomyces; alpha glucosidase; carbohydrate metabolism; chimeric proteins; enzyme induction /repression; enzyme structure; fermentation; fungal genetics; galactokinase; gene deletion mutation; genetic mapping; glucose; maltose; microorganism culture; nucleic acid hybridization; nucleic acid sequence; posttranslational modifications; protein biosynthesis; protein structure function; 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. We have chosen maltose fermentation in Saccharomyces as an example of a regulated system. We have chosen maltose fermentation in Saccharomyces as an example of a regulated system. The initial steps of maltose fermentation are carried out by maltose permease and maltase. Synthesis of these enzymes is induced by maltose and the Mal-activator mediates this regulation. The Ma1-activator is a DNA-binding transcription activator. We identified the functional domains of the Ma-1 activator, and localized the maltose- responsive regulatory domain to approximately the 220 residues at the C- terminus of this 470 residue protein. Our results indicate that several negative and positive regulatory sequence elements are found in the regulatory domain suggesting that several proteins interact with this region of Ma163p and regulate the transactivation domain of Ma163p in a sequence-specific manner. During the next grant period, we plan to test our model of maltose-dependent regulation of the Mal-activator. We will carry out a biochemical analysis of the Mal-activator protein and identify interacting proteins involved in maltose-regulated transcription activation. We will explore the regulatory role of subcellular localization, dimerization, formation of high molecular weight complexes, and post-translational modification in controlling DNA-binding and transcription activation by the Mal-activator. We will carry out a systematic mutation analysis of the regulatory domain of the Mal-activator to identify and localize the negative and positive regulatory sequence elements in this region. Allele-specific and high-copy suppressors of these mutant alleles will be isolated in an effort to identify proteins which interact with the Mal-activator at these sites. The Two-hybrid method will be used top identify proteins which interact with the Mal- activator. We will use molecular genetic approaches to determine whether maltose binds directly to the Ma1-activator. Mutations which broaden inducer specificity will be isolated. The alpha-methylglucoside-activator gene will be isolated and used to characterize the inducer sensing- signaling pathway.

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。我们有 糖疗法中选择的麦芽糖发酵为受调节的一个例子 系统。我们选择了糖疗法中的麦芽糖发酵为例 一个受管制的系统。麦芽糖发酵的最初步骤是 由麦芽糖渗透酶和麦芽糖酶进行。这些酶的合成是 由麦芽糖诱导，而麦芽活化剂介导了这种调节。这 MA1激活剂是DNA结合转录激活剂。我们确定了 MA-1激活剂的功能域，并将麦芽糖定位 响应性的调节域对C-的大约220个残基 该470个残基蛋白的末端。我们的结果表明有几个 在 调节域，表明几种蛋白质与此相互作用 MA163P的区域并调节A中MA163P的反式激活结构域 序列特异性的方式。在下一个赠款期间，我们计划测试 我们的麦芽糖依赖性调节模型。 我们将对MAL-激活蛋白的生化分析和 识别参与麦芽糖调节转录的相互作用蛋白 激活。我们将探讨亚细胞的调节作用 位置，二聚，高分子量复合物的形成， 以及控制DNA结合和的翻译后修改和 MAL激活剂的转录激活。我们将执行 MAL-ACRIVITOR的调节结构域的系统突变分析 识别和定位负调节序列 该地区的元素。等位基因特异性和高拷贝抑制剂 这些突变等位基因将被隔离以识别蛋白质 在这些位点与MAL-ACRIVITOR相互作用。两个杂交 方法将用于顶部识别与Mal-相互作用的蛋白质 激活剂。我们将使用分子遗传学方法来确定是否 麦芽糖直接与MA1激活剂结合。扩展的突变 诱导者特异性将被隔离。 α-甲基葡萄糖苷激活剂 基因将被分离，并用于表征诱导剂传感 - 信号通路。