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Transcriptional regulation of kidney-specific genes

肾脏特异性基因的转录调控

基本信息

批准号：
08457285
负责人：
INASE Naohiko
金额：
$ 5.25万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457285/
关键词：
AQP-2 AQP-3 CLC-K1 CLC-K2 Promoter CRE GATA purine-rich sequence 腎臓輸送体転写調節核内タンパク遺伝子制御

项目摘要

Expression of aquaporin-2 (AQP2) is exclusively limited to kidney collecting duct cells, and this strictly limited expression could be mediated by transcription of the gene. The 5-flanking region of the AQP2 gene was characterized by a TATA box, two GATA consensus sequences, an AP-1 site, an AP-2 site, three E-boxes, and a cycle AMP-responsive element. A reporter gene assay performed in the 1st day of primary culture of IMCD cells showed that the 5' region up to -2.9 kb worked as a promoter. Deletion experiments showed that at least two regions, from -434 to -364 and from -153 to -84, contain negatively acting cis-elements. To evaluate the functional role of GATA motifs in AQP2 gene, we sought to isolate a GATA factor(s) expressed in collecting ducts. Two cDNAs encoding GATA factors were isolated from rat kidney (GATA-2 and -3). GATA motifs in the 5-flanking region of the AQP2 gene were functional cis-elements and that GATA-3 in collecting ducts may be one of the important regulators o … More f AQP-2 expression in vivo.The rat CIC-K1 chloride channel is a kidney-specific member of the 010 chloride channel family found exclusively in the if-in ascending limb of Henle's loop in the kidney. To gain insight into the mechanism(s) of kidney-specific expression of CIC-K1, a genomic done that contains the 5'-flanking region of the rat CIC-K1 gene was isolated. The sequence of the proximal 5-flanking region contained an AP-3 site, a GRE, several AP-2 sites, and several E-boxes, but it lacked a TATA box.Deletion analysis revealed that this cell-specific promoter activity in IM cells was still present in the construct containing 51 b p of-the 5'-flanking region but was lost in the -29 construct, dearly demonstrating that the-22 bp from -51 to -30 have a major role in the cell-specific activity of the CIC-K1-promoter. These 22 bp consist of purine-rich sequence (GGGGAGGGG-GAGGGGAG), and gel-retardation analysis demonstrated the existence of a specific protein(s) binding to this element in IM cells. These results suggest that the novel purine-rich element may play a key role in the activity of the CIC-K1 gene promoter. Less

水通道蛋白-2（AQP 2）的表达仅限于肾集合管细胞，并且这种严格限制的表达可由该基因的转录介导。AQP 2基因的5-侧翼区的特征在于一个TATA盒、两个加塔共有序列、一个AP-1位点、一个AP-2位点、三个E盒和一个循环AMP响应元件。在IMCD细胞原代培养的第1天进行的报告基因测定表明，高达-2.9kb的5'区作为启动子起作用。缺失实验表明，至少有两个区域，从-434到-364和从-153到-84，含有负作用的顺式元件。为了评估加塔基序在AQP 2基因中的功能作用，我们试图分离在集合管中表达的加塔因子。从大鼠肾脏中分离出两个编码加塔因子的cDNA（加塔-2和-3）。AQP 2基因5-侧翼区的加塔基序是功能性顺式元件，集合管中的加塔-3可能是AQP 2基因表达的重要调控因子之一。 ...更多信息大鼠CIC-K1氯离子通道是010氯离子通道家族的肾特异性成员，仅在肾中亨利氏袢的if-in上升支中发现。为了深入了解CIC-K1在肾脏特异性表达的机制，分离了含有大鼠CIC-K1基因5 '侧翼区的基因组DNA。缺失分析显示，在IM细胞中，这种细胞特异性启动子活性在含有51个B p的5 '-侧翼区的构建体中仍然存在，但在-29构建体中丢失，清楚地表明-51至-30的-22bp在CIC-K1启动子的细胞特异性活性中具有重要作用。这22个碱基组成的嘌呤丰富的序列（GGGGAGGGG-GAGGGGAG），和凝胶阻滞分析表明存在一个特定的蛋白质结合到IM细胞中的这个元素。这些结果表明，新的富含嘌呤的元件可能在CIC-K1基因启动子的活性中起关键作用。少