AP-1 Complexes and Target Gene Regulation

AP-1 复合物和靶基因调控

• 批准号: 7468425
• 负责人: ELIZABETH J TAPAROWSKY
• 金额: $ 25.62万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7468425
• 关键词: Address; Affect; Affinity; Apoptosis; Binding; Biological; Blood Cells; Cell Differentiation process; Cells; Clinical; Complex; Coupled; DNA Binding; Development; Differentiation and Growth; Dimerization; Disease; Epitopes; Event; Exhibits; Extended Family; Family; Family member; Foundations; Gene Expression; Gene Expression Regulation; Gene Order; Gene Targeting; Genes; Genetic; Genetic Transcription; Genetically Engineered Mouse; Goals; Investigation; Knock-in Mouse; Knockout Mice; Laboratories; Link; Malignant Neoplasms; Mammalian Cell; Mediating; Modification; Molecular; Mus; Neurodegenerative Disorders; Numbers; Pattern; Phenotype; Phosphorylation; Positioning Attribute; Property; Proteins; Reagent; Regulation; Research; Role; Screening procedure; Signal Pathway; Signal Transduction; System; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic Intervention; Transactivation; Transcription Factor AP-1; Transgenes; bZIP Domain; cell growth; cell type; chromatin immunoprecipitation; design; hematopoietic tissue; human CREB3 protein; human disease; in vivo; information gathering; inhibitor/antagonist; killer T cell; leukemia; mouse model; novel; promoter; protein function; receptor; response; transcription factor

DESCRIPTION (provided by applicant): Intracellular signaling culminates in gene expression changes that are mediated by transcription factors such as activator protein-1 (AP-1). AP-1 is a widely expressed transcription complex consisting of dimerizing basic leucine zipper (bZIP) proteins. AP-1 functions in cell growth, differentiation, survival and apoptosis, and deregulated AP-1 activity figures prominently in human diseases from cancer to neurodegenerative disorders. Transcription factors are major targets for therapeutic intervention, and in vivo strategies designed to modulate AP-1 activity would be a significant advance in many clinical situations. BATF, and the highly related JDP1 protein, are unique AP-1 family members that dimerize with the central components of AP-1 - the Jun proteins - and inhibit AP-1 activity. The objective of this application is to define the roles of these AP-1 inhibitors in vivo and to test the hypothesis that these roles are linked to the regulation of specific AP-1 target genes. In Aim 1, genetically engineered mouse models expressing epitope-tagged BATF and JDP1 at endogenous levels will be used for defining the precise expression patterns of these proteins and for identifying genes directly bound by BATF using chromatin immunoprecipitation (ChIP) coupled with the screening of novel AP-1 promoter microarrays. BATF null, JDP1 null and double null mice will be used to assess how the loss of AP-1 inhibition mediated by these proteins affects development and to profile the full spectrum of gene expresson changes that characterize the BATF null phenotype. Aim 2 will use mass spectral analysis to identify and quantify discreet phosphorylation events that regulate the DNA binding and dimerization properties of BATF and influence its efficacy as an AP-1 inhibitor. Lastly, in Aim 3, the M1 cell system will be used to further investigate the function of BATF by following BATF-mediated regulation of target genes and by positioning BATF within the signaling network that controls the differentiation of these cells. These studies have the short-term goal of establishing the function of BATF and JDP1 in vivo and will generate the information necessary to address our long-term goals of using these AP-1 inhibitors for transcription-targeted therapies. Relevance: The aberrant cell growth that is a feature of many human diseases often is linked to deregulated gene expression mediated by proteins such as the AP-1 family of transcription factors. BATF and JDP1 have been identified as AP-1 family members that negatively regulate gene expression. The goal of this research is to fully characterize the biological activities of these proteins as a first step towards establishing their utility in AP-1 targeted therapies.

描述（由申请人提供）：细胞内信号传导最终导致由转录因子（例如激活蛋白-1 (AP-1)）介导的基因表达变化。 AP-1 是一种广泛表达的转录复合物，由二聚化碱性亮氨酸拉链 (bZIP) 蛋白组成。 AP-1 在细胞生长、分化、存活和凋亡中发挥作用，并且 AP-1 活性失调在从癌症到神经退行性疾病等人类疾病中发挥着重要作用。转录因子是治疗干预的主要目标，旨在调节 AP-1 活性的体内策略将在许多临床情况下取得重大进展。 BATF 和高度相关的 JDP1 蛋白是独特的 AP-1 家族成员，它们与 AP-1 的核心成分（Jun 蛋白）形成二聚体，并抑制 AP-1 活性。本申请的目的是确定这些 AP-1 抑制剂在体内的作用，并测试这些作用与特定 AP-1 靶基因的调节相关的假设。在目标 1 中，在内源水平表达表位标记的 BATF 和 JDP1 的基因工程小鼠模型将用于定义这些蛋白质的精确表达模式，并使用染色质免疫沉淀 (ChIP) 结合新型 AP-1 启动子微阵列的筛选来识别 BATF 直接结合的基因。 BATF null、JDP1 null 和双 null 小鼠将用于评估这些蛋白质介导的 AP-1 抑制的丧失如何影响发育，并分析表征 BATF null 表型的全谱基因表达子变化。目标 2 将使用质谱分析来识别和量化谨慎的磷酸化事件，这些事件调节 BATF 的 DNA 结合和二聚化特性并影响其作为 AP-1 抑制剂的功效。最后，在目标 3 中，M1 细胞系统将用于通过跟踪 BATF 介导的靶基因调节以及将 BATF 定位在控制这些细胞分化的信号网络内来进一步研究 BATF 的功能。这些研究的短期目标是确定 BATF 和 JDP1 的体内功能，并将产生必要的信息，以实现我们使用这些 AP-1 抑制剂进行转录靶向治疗的长期目标。 相关性：异常细胞生长是许多人类疾病的一个特征，通常与转录因子 AP-1 家族等蛋白质介导的基因表达失调有关。 BATF和JDP1已被鉴定为负调控基因表达的AP-1家族成员。这项研究的目标是全面表征这些蛋白质的生物活性，作为确定它们在 AP-1 靶向治疗中的效用的第一步。