TISSUE SPECIFIC CNTRL ALPHA 1B ANDRENOCEPTOR EXPRESSION

组织特异性 CNTRL ALPHA 1B 雄激素受体表达

基本信息

批准号：
6172833
负责人：
bin gao
金额：
$ 9.9万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-04-03 至 2000-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6172833
关键词：
DNA footprinting affinity chromatography alpha adrenergic receptor binding proteins cell type cofactor gel mobility shift assay genetic promoter element heart cell laboratory rat liver cells neoplastic cell receptor expression tissue /cell culture transcription factor transfection yeast two hybrid system

项目摘要

DESCRIPTION: The alpha1B adrenergic receptor (alpha1BAR) is a G-protein-coupled receptor that plays an important role in the control of cardiovascular homeostasis and metabolic processes in the liver, and recent studies have shown differential regulation of the transcription of the alpha1BAR gene in heart and liver under a variety of conditions. Dr. Gao will test the hypothesis that this differential regulation may be due to the tissue specific modulation of the action of the same transcription factor or to the interaction of the gene with different, tissue-specific transcription factors. Nuclear factor 1 (NF1) activates transcription driven by the dominant P2 promoter of the alpha1BAR gene in Hep3B cells and primary hepatocytes, but inhibits it in DDT1MF-2 cells. This duality may be due to modulation of the activity of NF1 by cell-specific cofactors. These cofactors will be identified by using the yeast two-hybrid system. The tissue distribution of NF1 parallels the relative abundance of the 2.7-kb alpha1BAR mRNA species, except for the heart, which expresses high levels of alpha1BAR and its 2.7 kb mRNA but little, if any, NFl. This suggests that transcription of the alpha1BAR gene in heart involves factor(s) other than NF1. Indeed, DNase I footprinting and DNA gel mobility shift analyses showed that the region -721 to -723 within the P2 promoter binds a new, previously unidentified transcription factor, called HF, in heart. This factor will be further characterized, purified by DNA affinity chromatography and cloned, if necessary. The role of this factor in transcription control will be identified by mutational and transfection analyses. Identification of the cell type-specific cofactors involved in modulating function of NF1 and the cardiac-specific transcription factors involved in the transcription of alpha1BAR gene in heart will shed light on the molecular mechanisms involved in the tissue specific regulation of alpha1BAR gene expression.

描述：α1b肾上腺素能受体（alpha1bar）是一个 G蛋白偶联受体在控制中起着重要作用肝脏中的心血管稳态和代谢过程，最近研究表明，调节的转录差异在各种疾病下，心脏和肝脏中的α1bar基因。高博士将检验以下假设，即这种差异调节可能是由于组织特异性调节相同转录因子的作用或基因与不同组织特异性转录的相互作用因素。核因子1（NF1）激活了由 HEP3B细胞中α1BAR基因的主要P2启动子和原发性的P2启动子肝细胞，但在DDT1MF-2细胞中抑制它。这种二元性可能是由于通过细胞特异性辅助因子对NF1活性的调节。这些将使用酵母双杂交系统来识别辅因子。这 NF1的组织分布与2.7-kb的相对丰度相似 α1barmRNA物种，除了心脏，它表达高水平 Alpha1bar及其2.7 KB mRNA，但很少（如果有）NFL。这表明这一点心脏中α1bar基因的转录涉及因素以外的因素 NF1。确实，DNase I足迹和DNA凝胶迁移率分析表明P2启动子内的区域-721至-723结合了一个新的，以前未识别的转录因子，称为HF，心中。这因素将进一步表征，通过DNA亲和力纯化必要时色谱和克隆。这个因素在转录控制将通过突变和转染确定分析。识别与细胞类型特异性辅助因子有关 NF1和心脏特异性转录因子的调节功能参与心脏中α1bar基因的转录会揭示参与组织特异性调节的分子机制 α1bar基因表达。