Abstract
To investigate the mechanisms of bradykinin B1 (BKB1) receptor gene expression, transient DNA transfection analyses of human BKB1 receptor gene promoter were performed in SV-40 transformed IMR90 cells. A positive regulatory element (PRE) located at position -604 to -448 base pair (bp) upstream of the transcription start site consistently exhibited, by far, the highest level of relative luciferase activity. A negative regulatory element, at position -682 to -604 bp, was able to completely ablate the function of the PRE. Transfection combined with deletion and mutation analyses illustrated that the PRE contains a classic, powerful enhancer. This enhancer was minimized to a 100-bp element at position -548 to -448 bp. A 78-bp fragment of negative regulatory element functioned as a silencer. Transient transfection of the enhancer construct, driven by heterologous herpes simplex thymidine kinase promoter, into a variety of cell types, showed that this enhancer presents a cell-type specific feature. In the characterization of the enhancer, motifs A (-548 to -532) and B (-483 to -477) were found to be essential for full enhancer activity. Motif D (-472 to -467) played a smaller role in enhancer activation. Gel shift and antibody supers...Continue Reading
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