Combinatorial Control of Transcription in Yeast

酵母转录的组合控制

• 批准号: 6624355
• 负责人: ANDREW VERSHON
• 金额: $ 28.7万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6624355
• 关键词: DNA binding protein; DNA footprinting; Saccharomyces cerevisiae; aminoacid metabolism; arginine; binding sites; cell growth regulation; crosslink; fungal genetics; gel mobility shift assay; gene expression; gene induction /repression; genetic promoter element; genetic regulation; genetic transcription; immunoprecipitation; mass spectrometry; mutant; protein protein interaction; protein structure function; site directed mutagenesis; transcription factor

DESCRIPTION (provided by applicant): Gene expression in eukaryotes is often under the control of multiple nuclear proteins that work in combination to regulate transcription. The combined interactions between these proteins often create a new form of regulatory activity that is distinct from either protein alone. The interactions between proteins dictate which DNA target sites are bound and therefore which sets of genes are regulated. In some cases, these protein interactions also influence whether a regulatory protein functions as a transcriptional activator or repressor. This form of regulation, often referred to as the combinatorial control of transcription, is widely used to control gene expression under specific cellular or developmental conditions. Understanding the mechanism of how these proteins interact in different combinations, and how these interactions determine their activity and specificity of the complex will provide insight into the regulation of many developmental and cellular processes. We have chosen two regulatory systems in the yeast Saccharomyces cerevisiae to investigate the mechanism of combinatorial control of transcription. The alpha2 repressor, a homeodomain protein, interacts with the Mcm1 and a1 protein to turn off two different sets of cell type specific-genes in yeast. We have previously investigated how alpha2 recognizes the DNA on its own, and how the interactions with Mcml and a1 influence the target specificity of the complex. We now propose to investigate how the a1-alpha2 complex binding to multiple sites in the HO promoter function to repress transcription of the gene (Specific aim 1). The HO gene contains a large and complex promoter with a wide array of different regulatory sites and serves as a good model system for the regulation of developmental genes in higher eukaryotes. The yeast Mcm1 protein is a transcriptional regulatory factor with sequence similarity to the MADS-box DNA-binding domains of the mammalian serum response factor (SRF) and Myocyte Enhancer Factor 2A (MEF2A) proteins. The MADS-box domain of Mcm1 interacts with at least five different cofactors to regulate different sets of genes that are required for cellular processes ranging from cell mating type and arginine metabolism to cell-cycle control. We propose to investigate how Mcm1 interacts with the MATalpha1 protein and how this interaction alters the conformation of the protein to activate transcription (Specific aim 2). We propose to investigate the interactions of Mcm1 with SFF, a complex of proteins including the forkhead protein, Fkh2, that regulate cell-cycle specific genes (Specific aim 3). Finally, we plan to investigate the interactions of Mcm1 with ArgR, a complex of three proteins that regulate genes involved in arginine metabolism. The information we learn about the interactions between these different classes of proteins will be relevant to more complex systems in higher eukaryotes and will provide insight on how these proteins function to regulate development.

描述（由申请人提供）：真核生物中的基因表达通常是 在多种核蛋白的控制下，结合起作用 调节转录。这些蛋白质之间的结合相互作用经常 创建一种与任何一种蛋白质不同的调节活动形式 独自的。蛋白质之间的相互作用决定了哪些DNA靶位点是 绑定，因此调节了哪一组基因。在某些情况下，这些 蛋白质相互作用还会影响调节蛋白是否起作用 转录激活器或阻遏物。这种法规形式，经常被称为 作为转录的组合控制，广泛用于控制 基因表达在特定的细胞或发育条件下。 了解这些蛋白质如何在不同的相互作用的机制 组合以及这些相互作用如何决定它们的活动和 综合体的特异性将为许多人的调节提供深入的了解 发育和细胞过程。我们选择了两个监管系统 酿酒酵母研究的酵母菌 转录组合控制。 Alpha2抑制剂（一种同源域蛋白）与MCM1和A1相互作用 蛋白质以酵母中关闭两种不同的细胞类型特异性基因。我们 以前已经研究了Alpha2如何单独识别DNA，以及如何 与MCML和A1的相互作用影响了目标特异性 复杂的。现在，我们建议研究A1-Alpha2复合物如何与 HO启动子功能中的多个位点抑制基因的转录 （特定目标1）。 HO基因包含一个宽大的大型启动子 一系列不同的监管站点，并用作良好的模型系统 高等真核生物中发育基因的调节。 酵母MCM1蛋白是带有序列的转录调节因子 与哺乳动物血清反应的MADS-Box DNA结合域相似 因子（SRF）和肌细胞增强子系数2A（MEF2A）蛋白。 Mads-Box MCM1的域与至少五个不同的辅助因子相互作用以调节 细胞过程所需的不同基因范围从 细胞交配类型和精氨酸代谢对细胞周期控制。我们建议 研究MCM1如何与Matalpha1蛋白相互作用以及如何相互作用 相互作用改变蛋白质的构象以激活转录 （特定目标2）。我们建议研究MCM1与SFF的相互作用， 一种调节的蛋白质的蛋白质，包括叉子蛋白FKH2 细胞周期特异性基因（特定目标3）。最后，我们计划调查 MCM1与ARGR的相互作用，ARGR是调节基因的三种蛋白质的复合物 参与精氨酸代谢。我们了解的信息 这些不同类别的蛋白质之间的相互作用将与 更高的真核生物中更复杂的系统，将提供有关这些方式的见解 蛋白质功能以调节发育。