TRANSCRIPTION COFACTORS OF THE THYROID HORMONE RECEPTOR

甲状腺激素受体的转录辅助因子

• 批准号: 6489709
• 负责人: JOSEPH D FONDELL
• 金额: $ 15.24万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-15 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6489709
• 关键词: cofactor; gel mobility shift assay; gene deletion mutation; genetic promoter element; genetic transcription; hormone receptor; molecular cloning; protein protein interaction; protein structure function; site directed mutagenesis; thyroid hormones; transcription factor; triiodothyronine

DESCRIPTION (Adapted from the applicant's abstract): The long-term stated goal of this proposal is to understand how auxiliary factors modulate transcriptional regulation by the thyroid hormone receptor (TR). This investigator has isolated a distinct group of novel proteins termed TRAPs (thyroid receptor associated proteins). In vitro transcription assays showed that the TR-TRAP complex markedly activates transcription from promoter templates containing TR-binding sites. He has cloned one of the TRAPs (TRAP 220) and found it directly interacts with TR in vivo in a T3-dependent manner. This protein shares significant sequence homology with PBP, a putative co-activator of peroxisome proliferator-activated receptors (PPARs), and RB18A, a novel p53-like co-regulatory protein. The investigator hypothesizes that TRAP 220, alone or in concert with other TRAP subunits, functions as a positive co-activator for gene-specific transcription. The aims of the proposal are: (1) Characterize TRAP 220 function in terms of ligand- induced interactions with TR and other nuclear hormone receptors (NRs) using deletion and site-directed mutagenesis to define interaction motifs of both TRAP 220 and other NRs using various protein-protein interaction studies and DNA mobility shift assays. (2) Examine the functional relevance of TRAP 220 as a transcriptional co-activator for hormone-dependent activation by TR and other NRs in co-transfection assays and in vitro transcription analyses. (3) Identify the specific cofactors which physically and functionally interact with TRAP 220. He proposes to screen the TR-TRAP complex by far western analysis using TRAP 220 as a probe to identify TRAP 220-interacting factors and a panel of recently cloned TRAP subunits will be screened for TRAP 220 interactions using protein-protein interaction assays. Identified cofactors will be further assayed for in vivo and functional associations with TRAP 220.

描述（改编自申请人的摘要）：长期 该提案的既定目标是了解辅助因素如何 通过甲状腺激素受体调节转录调控 （TR）。这位研究人员分离出了一组独特的新型蛋白质 称为 TRAP（甲状腺受体相关蛋白）。体外 转录测定表明 TR-TRAP 复合物显着激活 从含有 TR 结合位点的启动子模板转录。他 克隆了一种 TRAP (TRAP 220) 并发现它直接相互作用 体内TR以T3依赖的方式。这种蛋白质共享 与 PBP 具有显着的序列同源性，PBP 是一种推定的共激活剂 过氧化物酶体增殖物激活受体 (PPAR) 和 RB18A p53 样共调节蛋白。研究者推测 TRAP 220，单独或与其他TRAP亚基一起，充当 基因特异性转录的正辅激活因子。该组织的目标 建议是： (1) 根据配体描述 TRAP 220 功能 诱导与 TR 和其他核激素受体 (NR) 的相互作用 使用删除和定点突变来定义相互作用 使用各种蛋白质-蛋白质的 TRAP 220 和其他 NR 的基序 相互作用研究和 DNA 迁移率变化测定。 (2) 检查 TRAP 220 作为转录共激活剂的功能相关性 共转染中 TR 和其他 NR 的激素依赖性激活 测定和体外转录分析。 (3) 明确具体的 与 TRAP 220 发生物理和功能相互作用的辅助因子。他 提议通过远期西方分析使用筛选 TR-TRAP 复合物 TRAP 220 作为探针来识别 TRAP 220 相互作用因子和面板 最近克隆的 TRAP 亚基将被筛选出 TRAP 220 使用蛋白质-蛋白质相互作用测定进行相互作用。已鉴定 将进一步检测辅助因子的体内和功能 与 TRAP 220 的关联。