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作用机制。 监管。选择研究的系统是酵母调控途径。 由基因CCR4、CRE1和CRE2定义。CCR4是必需的 葡萄糖耐受酒精脱氢酶(ADH2)等的转录 非发酵基因。CRE1和CRE2是抑制的负效应因子 CCR4或其他需要CCR4才能运行的因素。Cre基因 是SPT基因家族的两个成员的等位基因，其中突变 抑制HIS4和LYS2基因座上的启动子插入。CCR4基因是 也是SPT介导的基因表达所必需的。小组委员会的一些成员 基因家族已被鉴定为组蛋白和一个与TATA结合的基因 蛋白。Cre/SPT基因作为几种致病因子调控基因的鉴定 显然不相关的系统，作为染色质和 转录复合体表明，这些调控因子与CCR4一起， 在维持基因的一般转录控制方面发挥关键作用 表情。 CCR4/Cre基因的分子作用将通过组合研究 遗传、生化和重组DNA技术。功能界别 CCR4和CRE1蛋白的区域将通过缺失和 突变分析，并将鉴定与CCR4结合的蛋白质 通过免疫学和生化技术。转录后 Cre基因对CCR4的调节将通过研究 CCR4与CRE1和CRE2的潜在蛋白质-蛋白质相互作用 确定cre突变对CCR4蛋白丰度的影响。这个 CCR4和cre突变对ADH2染色质结构的影响 以及这些因素在ADH2的作用地点 确认身份。还将确定CCR4功能所需的其他基因 通过进行额外的突变体搜寻，以及其他SPT的潜在作用 ADH2调控的基因将被检测。