Promoter Characterization of the Rat Gene for Ca2+-binding Protein Regucalcin
Promoter Characterization of the Rat Gene for Ca2+-binding Protein Regucalcin
复制标题
Ca2 结合蛋白 Regcalcin 的大鼠基因启动子特征
DOI：
10.1074/jbc.274.3.1277
复制
发表时间：
1999
期刊：
影响因子：
--
通讯作者：
M. Yamaguchi
中科院分区：
文献类型：
--
作者：
T. Murata;M. Yamaguchi
为了了解调节钙结合蛋白regucalcin基因表达的机制，我们对大鼠regucalcin基因的5′-侧翼区进行了表征。以大鼠肝帽位点cDNA为模板，用帽位点狩猎法确定了regucalcin基因的转录起始位点。将大鼠regucalcin基因5′侧翼区与荧光素酶报告基因连接后，在大鼠H4-II-E肝癌细胞中具有功能性启动子活性。3′-和5′-缺失分析表明大鼠regucalcin基因的基本功能启动子活性所需的序列。大鼠regucalcin基因的启动子活性增强与海湾K 8644，二丁酰cAMP，佛波酯，胰岛素和地塞米松治疗。利用凝胶迁移率变动分析，我们发现来自H4-II-E细胞的核蛋白特异性地结合到大鼠regucalcin基因的5′-侧翼区。此外，凝胶迁移率变动分析表明，Bay K 8644，二丁酰cAMP，佛波醇酯，胰岛素刺激核因子的结合到大鼠regucalcin基因的5′-侧翼区在H4-II-E细胞。这些结果表明，海湾K 8644-，二丁酰cAMP-，佛波酯-，和胰岛素诱导的核因子介导的刺激作用，每个调节器对大鼠regucalcin基因的启动子活性。
To understand the mechanism underlying the regulation of the Ca2+-binding protein regucalcin gene expression, we characterized the 5′-flanking region of the rat regucalcin gene. The transcriptional start site of the rat regucalcin gene was determined by the cap site hunting method with rat liver cap site cDNA. The 5′-flanking region of the rat regucalcin gene ligated to a luciferase reporter gene possessed functional promoter activity in rat H4-II-E hepatoma cells. 3′- and 5′-deletion analyses indicated the sequence required for basal functional promoter activity of the rat regucalcin gene. The promoter activity of the rat regucalcin gene was enhanced by treatment with Bay K 8644, dibutyryl cAMP, phorbol esters, insulin, and dexamethasone. Using gel mobility shift assays, we found that nuclear proteins from H4-II-E cells specifically bind to the 5′-flanking region of the rat regucalcin gene. Moreover, gel mobility shift assays revealed that Bay K 8644, dibutyryl cAMP, phorbol esters, and insulin stimulated the binding of nuclear factors to the 5′-flanking region of the rat regucalcin gene in H4-II-E cells. These results suggest that Bay K 8644-, dibutyryl cAMP-, phorbol ester-, and insulin-inducible nuclear factors mediate the stimulatory effect of each regulator on promoter activity of the rat regucalcin gene.
胰岛素刺激培养脂肪细胞中 c-Jun、c-Fos 和 Fos 相关蛋白的磷酸化。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kim,SJ;Kahn,CR
通讯作者：
Kahn,CR