HIGH MOBILITY GROUP PROTEIN IN GENE TRANSCRIPTION

基因转录中的高迁移率蛋白组

• 批准号: 2459700
• 负责人: Dimitris Thanos
• 金额: $ 24.55万
• 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/2459700
• 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; intermolecular interaction; nonhistone nucleoprotein; nuclear factor kappa beta; phosphorylation; 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（改编自申请人摘要）：真核生物 使用各种重要但知之甚少的机制来控制 他们基因的表达。 了解这些机制是一个 由于基因表达调控在 在一个细胞中，功能不同的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信号的反应能力 应力 这项研究计划的长远目标是 了解诱导基因表达的分子机制， 使用干扰素-β（IFN-β）基因作为模型系统的真核生物。 的 IFN-β基因通常被严格抑制，但在高水平表达。 当细胞被病毒感染或用双链 核糖核酸 以前的研究揭示了高流动性群体的关键作用 蛋白HMG I（Y）在IFN-β基因的病毒诱导中的作用。 HMG I（Y）不是一个 转录激活因子本身，但与其他 转录因子 IFN-β基因的高度特异性激活 为了应对病毒感染， 核蛋白转录增强子复合物，增强体。 HMG I 通过介导增强体的组装来发挥作用 蛋白质-DNA和蛋白质-蛋白质相互作用。 本项目的总体目标 本研究旨在阐明HMG I（Y） 促进增强体的组装以及增强体如何 协同地刺激转录。 首先，对HMG I（Y）进行了详细的结构和功能表征， 将进行蛋白质鉴定以鉴定涉及DNA结合的结构域， 蛋白质-蛋白质相互作用和组装所需的区域， IFN-β增强体的功能。 第二，体内试验和体外试验 转录系统将被用来确定的机制， IFN-β基因的组分协同增强了 转录激活。 如果成功，拟议的实验将 提供了关于结构和功能的重要新信息， 转录调控蛋白和序列，以及机制 参与信号传导。