YEAST GENES INVOLVED IN GENERAL TRANSCRIPTIONAL CONTROL

参与一般转录控制的酵母基因

• 批准号: 2180718
• 负责人: CLYDE L DENIS
• 金额: $ 21.27万
• 依托单位: UNIVERSITY OF NEW HAMPSHIRE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2180718
• 关键词: DNA binding protein; Saccharomyces cerevisiae; alcohol dehydrogenase; alleles; antibody; binding proteins; caseins; cell growth regulation; chimeric proteins; chromatin; fungal genetics; gene deletion mutation; gene expression; gene induction /repression; genetic promoter element; genetic regulatory element; genetic transcription; glutamine; histones; immunofluorescence technique; immunoprecipitation; laboratory rabbit; lethal genes; leucine; molecular cloning; nonhistone nucleoprotein; nucleic acid sequence; nucleosomes; phosphotransferases; plasmids; posttranslational modifications; protein structure function; recombinant DNA; site directed mutagenesis; temperature sensitive mutant; transcription factor; transfection

The goal of this research is to elucidate the mechanisms of eucaryotic gene regulation. The system chosen for study is the yeast regulatory pathway defined by the genes CCR4, CRE1, and CRE2. CCR4 is required for the transcription of glucose-repressible alcohol dehydrogenase (ADH2) and other non-fermentative genes. CRE1 and CRE2 are negative effectors that repress CCR4 or some other factor that requires CCR4 for function. The CRE genes are allelic to two members of the SPT gene family, mutations in which suppress promoter insertions at the HIS4 and LYS2 loci. The CCR4 gene is also required for spt-mediated gene expression. Some members of the SPT gene family have been identified as histones and one as the TATA binding protein. The identification of the CRE/SPT genes as regulators of several apparently unrelated systems and as integral components of chromatin and transcription complexes suggests that these regulators, together with CCR4, play key roles in maintaining general transcriptional control of gene expression. The molecular role of the CCR4/CRE genes will be studied by a combination of genetic, biochemical, and recombinant DNA techniques. The functional regions of CCR4 and CRE1 proteins will be identified by deletion and mutation analysis, and the proteins which bind to CCR4 will be identified by immunological and biochemical techniques. The post-transcriptional regulation of CCR4 by the CRE genes will be explored by examining the potential protein-protein interaction of CCR4 with CRE1 and CRE2 and by determining the effect of cre mutations on CCR4 protein abundance. The effects of ccr4 and cre mutations on ADH2 chromatin structure will be determined and the site at ADH2 through which these factors act will be identified. Other genes required for CCR4 function will also be identified by conduction additional mutant hunts, and the potential role of other SPT genes in ADH2 regulation will be examined.

本研究的目的是阐明真核基因的机制 规定。 选择用于研究的系统是酵母调控途径 由基因 CCR4、CRE1 和 CRE2 定义。 需要 CCR4 葡萄糖抑制性乙醇脱氢酶 (ADH2) 和其他酶的转录 非发酵基因。 CRE1 和 CRE2 是抑制的负效应子 CCR4 或需要 CCR4 发挥作用的其他一些因子。 CRE基因 与 SPT 基因家族的两个成员等位，其中的突变 抑制 HIS4 和 LYS2 位点的启动子插入。 CCR4基因是 spt 介导的基因表达也是必需的。 社会党的部分成员 基因家族已被确定为组蛋白，其中一个基因家族为 TATA 结合 蛋白质。 CRE/SPT 基因作为多种调节因子的鉴定 显然不相关的系统，作为染色质的组成部分和 转录复合物表明这些调节因子与 CCR4 一起， 在维持基因的一般转录控制中发挥关键作用 表达。 CCR4/CRE 基因的分子作用将通过组合进行研究 遗传、生物化学和重组 DNA 技术。 功能性的 CCR4 和 CRE1 蛋白的区域将通过删除和 突变分析，并鉴定与 CCR4 结合的蛋白质 通过免疫学和生化技术。 转录后 将通过检查 CRE 基因对 CCR4 的调节来探索 CCR4 与 CRE1 和 CRE2 之间潜在的蛋白质相互作用 确定 cre 突变对 CCR4 蛋白丰度的影响。 这 ccr4 和 cre 突变对 ADH2 染色质结构的影响 确定这些因素作用的 ADH2 位点 确定。 CCR4功能所需的其他基因也将被鉴定 通过进行额外的突变体狩猎，以及其他 SPT 的潜在作用 将检查 ADH2 调节中的基因。