Regulation and Function of Yeast Transcriptional Activator ADR1

酵母转录激活因子ADR1的调控和功能

• 批准号: 9513412
• 负责人: Clyde Denis
• 金额: $ 27.1万
• 依托单位: University of New Hampshire
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9513412&HistoricalAwards=false
• 关键词: Regulation; Function; Yeast; Transcriptional; Activator

9513412 Denis The goal of this research is to elucidate the mechanisms of eucaryotic gene regulation. The system chosen for study is the transcriptional activator ADR1 in its control of gene expression in the yeast Saccharomyes cerevisiae. The ADR1 protein contains four transactivation domains (TADs) and at least one inhibitory region (residues 227-229). The function of the TADs has been found to be highly dependent on the ADA2 coactivator complex. Moreover, ADA2 components interact specifically with individual ADR1 TADS, suggesting the ADA2 complex is a coactivator for ADRl. In addition, the 227-239 region of ADRl has been implicated in binding a repressor protein that inhibits ADR1 function independent of the promoter context. The central focus of this grant is to determine hmw the ADR1 activation and inhibitory domains interact with coactivators and repressors in bringing about the control of ADR1 function. The physical association of the ADA2 complex with ADRl will be explored biochemically, and the regions of the TADs required for binding this coactivator complex will be identified. The regions and residues of the ADRl TADs required for transactivation will be determined and correlated to those regions required for binding to the ADA2 complex. Using several complementary approaches, the putative repressor that binds the ADR1 inhibitory region will be cloned and characterized. These studies will help elucidate the complex interplay of a number of factors that can be involved in the control and action of transcriptional activators. %%% The focus of this project is a protein activator, ADRl, which stimulates the gene expression of certain proteins in the yeast Saccharomyces cerevisiae. ADRl binds to the DNA next to genes that produce enzymes that allow yeast to metabolize certain compounds when they are growing in air. The investigator has shown that ADR1 stimulates the production of RNA messages that produce the enzymes. ADR1 is a large protein containing four unique amino aci d segments that are required for the activation activity. His laboratory has found these segments must bind to certain other proteins to produce activation of gene expression. These other protein "coactivators" are a complex consisting of three proteins, ADA2/ADA3/GCN5. Proteins ADA2 and GCN5 specifically touch the sequences mentioned above. Another segment of ADR1 binds another protein that inhibits ADR1 activity. The investigator will characterize the contact areas between ADR1 and other proteins that are involved in regulating gene expression by ADR1. ***

本研究的目的是阐明真核生物基因调控的机制。本研究选择的系统是转录激活剂ADR1对酵母Saccharomyes酿酒酵母基因表达的控制。ADR1蛋白含有四个转激活结构域（tad）和至少一个抑制区（残基227-229）。已经发现TADs的功能高度依赖于ADA2共激活物复合物。此外，ADA2组分特异性地与ADR1 TADS相互作用，表明ADA2复合体是ADRl的辅激活因子。此外，ADRl的227-239区域与一种抑制蛋白的结合有关，这种抑制蛋白可以独立于启动子环境抑制ADR1的功能。这项资助的中心重点是确定ADR1激活和抑制结构域如何与共激活因子和抑制因子相互作用，从而控制ADR1功能。ADA2复合体与ADRl的物理关联将在生物化学上进行探索，并确定结合该共激活物复合体所需的TADs区域。ADRl TADs的交换激活所需的区域和残基将被确定，并与与ADA2复合物结合所需的区域相关。使用几种互补的方法，结合ADR1抑制区域的推定抑制因子将被克隆和表征。这些研究将有助于阐明可能参与转录激活因子的控制和作用的许多因素的复杂相互作用。这个项目的重点是一种蛋白质激活剂，ADRl，它能刺激酵母中某些蛋白质的基因表达。当酵母在空气中生长时，ADRl与产生酵素的基因旁边的DNA结合，酵素使酵母能够代谢某些化合物。研究者已经证明，ADR1刺激产生产生酶的RNA信息的产生。ADR1是一个大的蛋白质，包含四个独特的氨基酸d片段，这是激活活性所必需的。他的实验室发现，这些片段必须与某些其他蛋白质结合，才能激活基因表达。这些其他蛋白质“共激活因子”是由三种蛋白质组成的复合物，ADA2/ADA3/GCN5。蛋白质ADA2和GCN5特异性接触上述序列。ADR1的另一段结合另一种抑制ADR1活性的蛋白质。研究者将描述ADR1与其他参与调节ADR1基因表达的蛋白之间的接触区域。＊＊＊