TRANSCRIPTION COFACTORS OF THE THYROID HORMONE RECEPTOR

描述(改编自申请人的摘要)：长期 这项建议的既定目标是了解辅助因素如何 甲状腺激素受体对转录的调节作用 (Tr)。这位研究人员分离出了一组不同的新蛋白质 称为TRAP(甲状腺受体相关蛋白)。离体 转录分析表明，tr-trap复合体显著激活 从含有TR结合位点的启动子模板转录而来。他 克隆了其中一个陷阱(陷阱220)，并发现它直接相互作用 在体内以一种T3依赖的方式与tr结合。这种蛋白质与 与PBP的显著序列同源性，PBP可能是 过氧化物酶体增殖物激活受体(PPAR)和新的RB18A P53样共调控蛋白。调查员假设陷阱 220单独或与其他陷阱亚单位协同工作，起到 基因特异性转录的阳性共激活剂。该计划的目标是 建议如下：(1)从配体的角度表征TRAP 220的功能。 诱导与受体和其他核激素受体的相互作用 使用缺失和定点突变来定义相互作用 利用不同蛋白质-蛋白质的TRAP 220和其他近等基因的模体 相互作用研究和DNA迁移率改变分析。(2)审查 TRAP 220作为转录共激活因子的功能相关性 TRR和其他NRs在共转染中的激素依赖激活 检测和体外转录分析。(3)确定具体的 物理上和功能上与捕捉器220相互作用的辅因。他 建议筛选到目前为止的tr圈闭综合体使用西方分析 陷阱220作为识别陷阱220相互作用因素的探针和面板 将对最近克隆的TRAP亚基进行TRAP 220的筛查 使用蛋白质-蛋白质相互作用分析的相互作用。已确定 辅因子将进一步在体内和功能上进行检测 与陷阱220的关联。

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.