Regulation and Function of Yeast Transcriptional Activator ADR1

9513412 Denis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转录激活因子ADR 1在其控制的基因表达的酵母酿酒酵母。 ADR 1蛋白含有四个反式激活结构域（TADs）和至少一个抑制区（残基227-229）。 已发现TADs的功能高度依赖于ADA 2辅激活因子复合物。 此外，ADA 2组分与个体ADR 1 TADS特异性相互作用，表明ADA 2复合物是ADR 1的共活化剂。 此外，ADR 1的227-239区域与抑制ADR 1功能的阻遏蛋白结合有关，而不依赖于启动子环境。 这项研究的重点是确定ADR 1激活和抑制结构域与辅激活因子和抑制因子相互作用，从而控制ADR 1功能。 将以生物化学方式探索ADA 2复合物与ADR 1的物理缔合，并将鉴定结合该共激活因子复合物所需的TAD区域。 将确定反式激活所需的ADR 1 TAD的区域和残基，并将其与结合至ADA 2复合物所需的那些区域相关联。 使用几种互补的方法，结合ADR 1抑制区的推定阻遏物将被克隆和表征。这些研究将有助于阐明复杂的相互作用的一些因素，可以参与转录激活因子的控制和行动。 该项目的重点是一种蛋白质激活剂ADR 1，它可以刺激酿酒酵母中某些蛋白质的基因表达。 ADR 1与产生酶的基因旁边的DNA结合，这些酶允许酵母在空气中生长时代谢某些化合物。 研究人员已经表明，ADR 1刺激产生酶的RNA信息的产生。 ADR 1是一种大蛋白，含有四个独特的氨基酸片段，这些片段是激活活性所需的。 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，ADA 2/ADA 3/GCN 5。 蛋白质ADA 2和GCN 5特异性地接触上述序列。 ADR 1的另一个片段结合另一种抑制ADR 1活性的蛋白质。 研究者将表征ADR 1与其他参与ADR 1调节基因表达的蛋白质之间的接触面积。 ***

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. ***