Regulation and function of yeast transcr#itional activator ADR1

本研究的目的是阐明真核基因调控的机制。 选择用于研究的系统是酵母转录激活剂 ADR1，它控制葡萄糖抑制性乙醇脱氢酶 (ADH2)。 将特别强调定义ADR1的调节机制、ADR1的功能区域、介导或影响ADR1功能的因素，以及这些因素和ADR控制ADH2转录的方式。 将结合使用生化、重组 DNA 和遗传分析。 将鉴定 ADR1 的体内磷酸化位点，并确定 cAMP 依赖性 (cAPK) 和非依赖性蛋白激酶在 ADR1 调节中的作用。 ADR1 的识别机制。 将分离 ADR1 蛋白，并鉴定其对于转录激活、甘油生长调节、寡聚化、葡萄糖控制和娇小诱导的重要区域。 将对结合或介导 ADR1 作用的蛋白质进行生化和遗传学研究。 ADR1 激活 ADH2 转录所需的 SAF1-3 基因将被克隆并测序。 独立于 ADR1 控制 ADH2 发挥作用的 CCR1 和 SCH9 蛋白激酶的作用也将被确定。

The goal of this research is to elucidate the mechanisms of eucaryotic gene regulation. The system chosen for study is the yeast transcriptional activator ADR1 in its control of glucose repressible alcohol dehydrogenase (ADH2). Particular emphasis will be placed on defining the mechanisms by which ADR1 is regulated, the functional regions of ADR1, the factors which mediate or influence ADR1 function, and the means by which these factors and ADR control ADH2 transcription. A combination of biochemical, recombinant DNA, and genetic analyses will be used. The in vivo phosphorylation site of ADR1 will be identified and the role of cAMP-dependent (cAPK) and -independent protein kinases in ADR1 regulation will be ascertained. The mechanisms by which ADR1 will be identified. The ADR1 protein will be isolated and its regions important to transcriptional activation, regulation of glycerol growth, oligomerization, glucose control, and induction of petites will be identified. The proteins which bind to or mediate ADR1 action will be studied biochemically and genetically. The SAF1-3 genes required for ADR1 activation of ADH2 transcription will be cloned and sequenced. The role of the CCR1 and SCH9 protein kinases which act independently of ADR1 controlling ADH2 will also be determined.