权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The transcriptional regulation of the human β1-adrenergic receptor gene by stress-related transcription molecules

应激相关转录分子对人β1肾上腺素受体基因的转录调控

基本信息

批准号：
12670961
负责人：
MATSUI Hiroaki
金额：
$ 1.34万
依托单位：
St. Marianna University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

We characterized the transcription factors involved in the regulation of the human β1-adrenergic receptor (β1-AR) gene expression and studied effects of stress-related transcription molecules, such as glucocorticoid receptor(GR) and heat shock factors, on the transcription of the β1-AR gene in human neuroblastoma SY5Y ceils. Deletional and mutational analyses of the β1-AR gene promoter-luciferase reporter gene constructs revealed that the trimeric transcription factor NF-Y binding site, located in -369/-365 relative to the translation start codon, has a pivotal role in the regulation of the β 1-AR gene expression. In addition, we found one potential binding site for CREB (-353/-346) and two sites for AP-2 (-345/-337 and -332/-324) near the NF-Y binding site. By introducing mutations into either of the two AP-2 sites, the β1-AR gene expression was moderately reduced. In contrast, CREB binding site mutation was less effective in the reduction of the β1-AR gene expression. These results indicate that NF-Y regulates the β1-AR gene expression by recruiting AP-2. Treatment with glucocorticoid (GC) inhibited the β1-AR gene expression in SY5Y cells. Assuming that GR Interacts with NF-Y through the protein-protein interaction, GC-mediated inhibition of the β1-AR gene expression can occur through the sequestration of NF-Y by GR from the β1-AR gene promoter.

我们鉴定了参与调节人β1-肾上腺素能受体（β1-AR）基因表达的转录因子，并研究了应激相关转录分子，如糖皮质激素受体（GR）和热休克因子，对人神经母细胞瘤SY5Y细胞β1-AR基因转录的影响。β1-AR基因启动子荧光素酶报告基因构建的缺失和突变分析表明，三聚体转录因子NF-Y结合位点（相对于翻译起始密码子-369/-365）在β1-AR基因表达调控中起关键作用。此外，我们在NF-Y结合位点附近发现了CREB的一个潜在结合位点（-353/-346）和AP-2的两个潜在结合位点（-345/-337和-332/-324）。通过将突变引入两个AP-2位点中的任何一个，β1-AR基因的表达适度降低。相比之下，CREB结合位点突变在降低β1-AR基因表达方面效果较差。这些结果表明NF-Y通过募集AP-2调控β1-AR基因的表达。糖皮质激素（GC）抑制SY5Y细胞β1-AR基因表达。假设GR通过蛋白-蛋白相互作用与NF-Y相互作用，gc介导的β1-AR基因表达抑制可以通过GR从β1-AR基因启动子中分离NF-Y来实现。