EUKARYOTIC GENE REGULATION IN YEAST

酵母中的真核基因调控

• 批准号: 2176520
• 负责人: MARK S PTASHNE
• 金额: $ 34.79万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-07-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2176520
• 关键词: DNA binding protein; DNA directed RNA polymerase; Drosophilidae; Saccharomyces; eukaryote; fungal genetics; galactose; gene expression; genetic promoter element; genetic transcription; molecular cloning; mutant; nucleic acid sequence; protein structure; transcription factor

The long term goal of this grant is to further the understanding of how eukaryotic transcriptional regulators bind to DNA and affect gene transcription. Although the proposed experiments involve transcriptional regulators from a variety of organisms and address a variety of questions, all involve, at least in part, manipulations in yeast to facilitate achieving the goals. The specific aims fall into three broad categories. First, a series of experiments concern the mechanism by which GAL4, a widely studied "universal" activator from yeast, activates transcription. Particular attention will be paid to the effect of GAL11 an auxiliary factor that somehow helps GAL4 and certain other activators. Genetic and physiological experiments will probe the idea that GAL11 forms a complex with these activators, a complex that may involve additional proteins as well. Second, the mechanism by which members of the GAL4 family of activators recognize specific sites in DNA will be analyzed. This is a particularly intriguing example of protein DNA interaction: each of these proteins (eg GAL4, PUT3 and PPR1) bears similar "zinc cluster" elements that contact identical DNA triplets; the distinguishing feature of each DNA site is evidently the spacing between these recognized triplets. Among other experiments, chimeric proteins will be analyzed to reveal the basis of the protein DNA specificity. Third, the action of the Drosophila morphogen Dorsal will be analyzed. The putative co-factor that presumably converts Dorsal from an activator into a negative regulator will be cloned, as will the natural Drosophila activator that is inhibited by Dorsal working in conjunction with its putative co-factor. The structural basis for the interaction between Dorsal and Cactus, and between Dorsal (a member of the rel class of activators) and DNA will be analyzed.

这笔赠款的长期目标是进一步了解 真核转录调节剂与DNA结合并影响基因 转录。 尽管提出的实验涉及转录 来自各种生物的监管机构，解决了各种问题， 至少部分涉及酵母中的操作以促进 实现目标。 具体目标分为三个广泛的类别。 首先，一系列实验涉及gal4的机制 广泛研究了酵母的“通用”激活剂，激活转录。 特别关注GAL11辅助的效果 以某种方式帮助Gal4和某些其他激活剂的因素。 遗传和 生理实验将探究GAL11形成复合物的想法 使用这些激活剂，一种可能涉及其他蛋白质的复合物作为 出色地。 其次，GAL4家族成员的机制 激活剂将识别DNA中的特定位点。 这是一个 特别有趣的蛋白质DNA相互作用的例子：这些 蛋白质（例如Gal4，put3和ppr1）具有相似的“锌簇”元素 接触相同的DNA三胞胎；每个的区别特征 DNA位点显然是这些公认的三胞胎之间的间距。 除其他实验外，还将分析嵌合蛋白以揭示 蛋白DNA特异性的基础。 第三，果蝇的作用 将分析形态的背。 推定的联合因素大概是 将背面从激活器转换为负调节器 克隆，天然果蝇激活剂也被抑制 背侧与推定的共同因素一起工作。 结构 背侧和仙人掌之间以及背侧之间相互作用的基础（a 将分析REL类激活剂的成员）和DNA。