Negative control elements and cAMP responsive sequences in the tissue-specific expression of mouse renin genes.
Negative control elements and cAMP responsive sequences in the tissue-specific expression of mouse renin genes.
复制标题
小鼠肾素基因组织特异性表达中的阴性控制元件和 cAMP 响应序列。
DOI：
10.1073/pnas.86.1.56
复制
发表时间：
1989
影响因子：
11.1
通讯作者：
Dzau,VJ
中科院分区：
文献类型：
--
作者：
Nakamura,N;Burt,DW;Paul,M;Dzau,VJ
小鼠肾素基因的5 '侧翼区（Ren1d和Ren2d）含有推定的阴性对照和cAMP反应元件。序列分析还表明，这些假定的控制元件在Ren2d基因中断的160个碱基对插入。为了记录这些元件的功能，我们分离出这些区域，并将其与报告基因氯霉素乙酰转移酶（CAT）融合，该报告基因上游连接到胸苷激酶（TK）启动子（pUTKAT 1）。将嵌合体转染入小鼠垂体瘤AtT-20和人绒毛膜癌JEG-3细胞中。在基础未刺激条件下，Ren 1d 5 '侧翼序列在正义方向抑制pUTKAT 1的TK启动子的基础CAT表达，而相同序列在反义方向则没有。Ren2d的5 '侧翼区对基础CAT表达没有抑制作用。这些数据表明，阴性对照元件在Ren1d中有功能，但在Ren2d中无功能，表明Ren2d中的160个碱基对插入干扰了阴性对照元件的功能。在响应8-溴-cAMP，两个肾素基因增加转录3倍，这表明在两个基因的cAMP响应元件的功能顺式作用。这些数据可能是重要的，在了解小鼠肾素基因的组织特异性表达的调节。
The 5' flanking regions of the mouse renin genes (Ren1d and Ren2d) contain putative negative control and cAMP responsive elements. Sequence analysis shows additionally that these putative control elements in the Ren2d gene are interrupted by a 160-base-pair insertion. To document the functions of these elements, we isolated these regions and fused them to the reporter gene chloramphenicol acetyltransferase (CAT), which was linked upstream to a thymidine kinase (TK) promoter (pUTKAT1). The chimeric constructs were transfected into mouse pituitary tumor AtT-20 and human choriocarcinoma JEG-3 cells. At the basal unstimulated condition, Ren1d 5' flanking sequence in the sense orientation inhibited basal CAT expression from the TK promoter of pUTKAT1, whereas the same sequence in the antisense orientation did not. The 5' flanking region of Ren2d had no inhibitory effect on basal CAT expression. These data demonstrate that the negative control element is functional in Ren1d but is nonfunctional in Ren2d, suggesting that the 160-base-pair insertion in Ren2d interferes with the function of the negative control elements. In response to 8-bromo-cAMP, both renin genes increased transcription 3-fold, suggesting a functional cis action of the cAMP responsive element in both genes. These data may be important in the understanding of the regulation of the tissue-specific expression of mouse renin genes.