Abstract
Nested deletion mutants of the 5' flanking region of the β-cell transcription unit of the rat glucokinase gene (rßGK) were fused to the CAT-reporter gene. Transient expression studies in HIT M2.2.2 and BHK21 cells revealed a distal (upstream of −359) and a proximal promoter region (between −278/−49) harbouring positive and negative regulatory elements. DNaseI footprinting revealed three protected areas between nucleotides −190 and −60. DNA-elements playing a crucial role in transcriptional control of the insulin genes (IEB- and CT-motifs) have been detected within the proximal promoter region and contribute to β-cell specific gene regulation. 3' deletion analysis revealed that DNA-elements located downstream from transcription initiation sites (up to +123) contribute to transcriptional regulation.
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