We isolated a genomic clone containing 2558 bp of the 5′-flanking region and 217 bp of the first exon for the human type-1 angiotensin II receptor (AT1) gene. Primer extension and RNAase protection analyses identified a transcriptional start site (+1) at 39 and 114 bp, downstream of putative TATA and GC boxes, respectively. Chimeras containing 2.6 kbp(-2558 to +79) of the 5′-flanking region and chloramphenicol acetyltransferase(CAT) gene expressed a significant CAT activity when transfected into bovine aortic smooth muscle cells(BASMC), but not the cells which had no detectable AT1 receptors, such as HeLa cells and primary cultured human skin fibroblasts. Deletion of the 5′-flanking region up to position -114 resulted in more than 20-fold increase of the reporter activity in BASMC, suggesting the presence of negatively regulating element(s) in the upstream promoter region. These results indicate that we have cloned a functional promoter for the human AT1 receptor gene.
|Number of pages||7|
|Journal||Biochemical and Biophysical Research Communications|
|Publication status||Published - May 15 1994|
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology