Regulation of Transcription at the Human IL-2 Promoter

人 IL-2 启动子的转录调控

• 批准号: 7936648
• 负责人: James Goodrich
• 金额: $ 4.31万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936648
• 关键词: Activator Appliances; Affinity; Antigens; Binding; Biochemical; Biological Assay; Biological Process; Cells; Complex; Coupled; DNA; Data; Disease; Elements; Event; Genes; Genetic Transcription; Goals; Human; Immune response; Indium; Infection; Interleukin-2; Kinetics; Measures; Mediating; Messenger RNA; Methods; Molecular; Molecular Conformation; Molecular Genetics; Molecular Profiling; Mutation; Normal Cell; Nucleic Acid Regulatory Sequences; Nucleoproteins; Pattern; Peptides; Polymerase; Production; Property; Proteins; RNA Polymerase II; Recruitment Activity; Regulation; Research; Role; Signal Transduction; Signaling Molecule; Site; Structure; T-Cell Activation; T-Lymphocyte; TAF1 gene; TAF4 gene; Testing; Time; Transcription Coactivator; Transcriptional Activation; Transcriptional Regulation; Work; anergy; base; cell growth; combinatorial; cytokine; genome-wide; inhibitor/antagonist; innovation; insight; novel; programs; promoter; protein complex; protein protein interaction; research study; response; stoichiometry; transcription factor

DESCRIPTION: (provided by applicant): Interleukin-2 (IL-2) is a cytokine that functions in the mammalian immune response to infection. During an immune response, production of IL-2 is upregulated at the level of transcription by activators such as NFAT1 and cJun. The main goal of this research is to understand the mechanisms of transcriptional activation at the human IL-2 promoter. An additional goal is to reveal the extent to which three protein-protein interactions control transcription genome-wide when human T cells respond to both activating and anergic signals. The results of these studies will provide significant insight into how mRNA transcription is regulated, which is vitally important in understanding normal cell growth as well as aberrations associated with many diseases. The studies will decipher how promoter DNA elements, protein-protein interactions, and a synergistic nucleoprotein complex contribute to setting the level of IL-2 transcription in T cells under different stimulatory conditions. The specific aims of the proposal are the following: 1) To establish a kinetic profile of the molecular composition at the IL-2 promoter during co-stimulation and anergy, and determine whether promoter-proximal pausing is involved in IL-2 regulation. 2) To determine the role of three protein-protein interactions in setting the program of molecular events that occurs at the IL-2 promoter during T cell activation. 3) To obtain an understanding of how specific protein-protein interactions control the two different transcriptional programs that occur in co-stimulated and anergic T cells. 4) To investigate the mechanism by which transcriptional activators, distinct DNA elements, and nucleoprotein structure contribute to combinatorial synergistic activation at the IL-2 promoter. 5) To understand the biophysical properties of interactions between transcription factors by measuring the interaction affinities and subunit stoichiometries of complexes containing defined regions of specific activators and coactivators. These studies utilize both cell-based and biochemical assays. One of the innovative aspects of this research is the use of inhibitory peptides to study the function of specific protein-protein interactions between cJun, NFAT1, and two subunits of the TFIID complex in setting transcriptional programs in human T cells. The proposed experiments will reveal novel mechanisms of transcriptional regulation at the IL-2 promoter and other promoters in human T cells by providing general insight into: how promoter elements influence synergistic transcriptional activation, how protein-protein interactions contribute to transcriptional regulation genome-wide, and how nucleoprotein complexes form and function at the regulatory region of a natural human promoter.

描述：(申请人提供)：白介素2(IL-2)是一种在哺乳动物对感染的免疫反应中起作用的细胞因子。在免疫反应中，IL-2的产生被NFAT1和cJun等激活剂在转录水平上上调。本研究的主要目的是了解人IL-2启动子转录激活的机制。另一个目标是揭示当人类T细胞同时对激活和无能信号做出反应时，三种蛋白质-蛋白质相互作用在多大程度上控制整个基因组的转录。这些研究的结果将提供对mRNA转录如何调控的重要洞察，这对于理解正常的细胞生长以及与许多疾病相关的异常是至关重要的。这些研究将破译启动子DNA元件、蛋白质-蛋白质相互作用和协同核蛋白复合体如何在不同刺激条件下帮助设定T细胞中IL-2的转录水平。该提案的具体目的如下：1)建立共刺激和无能时IL-2启动子分子组成的动力学图谱，并确定启动子-近端停顿是否参与IL-2的调节。2)确定三种蛋白质相互作用在设定T细胞活化过程中IL-2启动子发生的分子事件的程序中的作用。3)了解特定的蛋白质-蛋白质相互作用如何控制共刺激和无能T细胞中发生的两种不同的转录程序。4)探讨转录激活剂、不同的DNA元件和核蛋白结构在IL-2启动子协同激活中的作用机制。5)通过测量包含特定激活剂和辅激活剂特定区域的复合体的相互作用亲和力和亚单位化学计量，了解转录因子之间相互作用的生物物理性质。这些研究同时利用了基于细胞的分析和生化分析。本研究的创新方面之一是使用抑制肽来研究cJun、NFAT1和TFIID复合体的两个亚单位之间的特定蛋白质-蛋白质相互作用在设定人类T细胞转录程序中的功能。拟议的实验将揭示在人类T细胞中IL-2启动子和其他启动子的转录调控的新机制，通过提供总体洞察：启动子元件如何影响协同转录激活，蛋白质-蛋白质相互作用如何在全基因组范围内促进转录调控，以及核蛋白复合体如何在自然人类启动子的调节区形成和功能。