HIGH MOBILITY GROUP PROTEIN IN GENE TRANSCRIPTION

基因转录中的高迁移率蛋白组

• 批准号: 6019171
• 负责人: Dimitris Thanos
• 金额: $ 25.95万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2001-07-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6019171
• 关键词: DNA binding protein; DNA footprinting; chimeric proteins; gel mobility shift assay; gene induction /repression; genetic enhancer element; genetic promoter element; genetic regulation; genetic transcription; immunoprecipitation; interferon beta; nonhistone nucleoprotein; nuclear factor kappa beta; phosphorylation; protein protein interaction; protein structure function; stoichiometry; transcription factor; western blottings

DESCRIPTION (Adapted from Applicant's Abstract): Eukaryotic organisms employ a variety of important, but poorly understood mechanisms to control the expression of their genes. An understanding of these mechanisms is an important issue since regulation of gene expression plays a pivotal role in the development and differentiation of functionally distinct cell types in a precise spatial manner. In addition, regulation of transcription reflects the ability of cells to respond to extracellular signals and environmental stress. The long term objectives of this research proposal are to understand the molecular mechanisms of inducible gene expression in higher eukaryotes using the interferon-beta (IFN-beta) gene as a model system. The IFN- beta gene is ordinarily tightly repressed, but is expressed at high levels when cells are infected with viruses or treated with double stranded RNA. Previous studies revealed a key role for the High Mobility Group protein HMG I(Y) in virus induction of the IFN-beta gene. HMG I(Y) is not a transcriptional activator on its own but functions in concert with other transcription factors. The highly specific activation of the IFN-beta gene in response to virus infection requires the assembly of a higher order nucleoprotein transcription enhancer complex, the enhanceosome. HMG I functions by promoting the assembly of the enhanceosome by mediating protein-DNA and protein-protein interactions. The overall goals of this proposal are to elucidate the molecular mechanisms by which HMG I(Y) promotes the assembly of the enhanceosome and how the enhanceosome stimulates transcription synergistically. First, a detailed structural and functional characterization of the HMG I(Y) proteins will be carried out to identify domains involved in DNA binding, protein-protein interactions and regions required for the assembly and the function of the IFN-beta enhanceosome. Second, in vivo assays and in vitro transcription systems will be used to determine the mechanisms by which the components of the IFN-beta gene enhanceosome function synergistically in the activation of transcription. If successful, the proposed experiments will provide significant new information regarding the structure and function of transcriptional regulatory proteins and sequences, as well as the mechanisms involved in signal transduction.

描述(摘自申请者摘要)：真核生物 使用各种重要但鲜为人知的机制来控制 他们基因的表达。对这些机制的理解是一个 重要问题，因为基因表达的调节在 一种不同功能细胞类型的发育和分化 精确的空间方式。此外，转录调控反映了 细胞对细胞外信号和环境做出反应的能力 压力。这项研究计划的长期目标是 了解高等植物诱导基因表达的分子机制 使用干扰素-β基因作为模型系统的真核生物。这个 干扰素-β基因通常被严格抑制，但在高水平表达 当细胞感染病毒或双链处理时的水平 核糖核酸。之前的研究表明，高流动性人群扮演着关键角色 HMG I(Y)蛋白在病毒诱导干扰素-β基因中的作用。HMG I(Y)不是 转录激活物本身，但与其他 转录因子。干扰素-β基因的高度特异性激活 为了应对病毒感染，需要更高层次的组装 核蛋白转录增强子复合体，增强体。HMG I 通过调节促进增强体的组装而发挥作用 蛋白质-DNA和蛋白质-蛋白质相互作用。这个项目的总体目标是 建议阐明HMG I(Y)的分子机制 促进增强体的组装以及增强体如何 协同刺激转录。 首先，详细描述了HMG I(Y)的结构和功能。 蛋白质将被用来识别与DNA结合有关的结构域， 蛋白质相互作用和组装所需的区域 干扰素-β增强体的功能。第二，体内检测和体外检测 转录系统将被用来确定 干扰素-β基因增强小体功能的组份在 转录激活。如果成功，拟议中的实验将 提供有关结构和功能的重要新信息 转录调控蛋白和序列及其作用机制 参与信号转导。