Identification of cis- and trans-acting factors regulating the expression of the human insulin receptor gene.

The functional organization of the human insulin receptor (hIR) promoter was analyzed by deletion mutagenesis and protein-DNA interaction studies. A series of deletion mutants was expressed transiently in two human hepatocytes, HepG2 and PLC. The results revealed that the promoter region between -692 and -345 is essential for efficient transcription of the hIR gene. Multiple trans-acting factors were identified by band shift and footprinting analyses. Sp1 binds to a cluster of GC boxes and two GGGAGG hexamers locating at -637 to -594. Adjacent to GC boxes, there are two regions, from -550 to -530 and from -522 to -503, which bind to two novel factors, IRNF-I and IRNF-II. These two factors are distributed differentially in different cell lines. Linker scanning mutations on GC, GA boxes, or the IRNF-I binding site significantly decreased the transcriptional activity, indicating that IRNF-I and Sp1 are important for hIR promoter activity. In addition, we demonstrated that glucocorticoid-dependent transcriptional induction of hIR mRNA in vivo is conferred by a glucocorticoid response element in the hIR promoter. Taken together, these results imply that transcription of the human insulin receptor gene is regulated by multiple protein-DNA interactions occurring within the defined promoter region.