Identification and characterization of a glucose-responsiveness region upstream of human insulin gene in transfected HIT-T 15 cells.
To determine possible regulation of full-length human insulin gene promoter activity by glucose, we examined a 2-kilobase pair (kbp) 5'-flanking region of the human insulin gene and characterized the DNA elements in transfected HIT-T 15 cells. The expression of the 2-kilobase pair 5'-flanking region human insulin gene fused to the luciferase reporter gene occurred by transfection. In 0.8 mM glucose of the F-12 K medium, the element mediating the negative regulatory region was localized from -1782 to -1295 base pairs (bp) and stimulatory element from -1295 to -1138 bp. The elements from -1138 to -880 bp and from -356 to +252 bp possessed the elements dose-dependently responsive to 0.8 mM, 7.0 and 22.2 mM glucose. In fragment D, cotransfection of oligonucleotide that confers RIPE3b1 activator decreased the glucose-stimulated promoter activity, but the other oligonucleotide that confers STF-1 did not. The present data indicated that 2 kbp possesses glucose-responsive region in the element from -1138 to -880 bp, in addition to the previously reported element from -356 to initiation site. There may exist a RIPE3b1 activator binding site in the glucose-responsive element from -1138 to -880 bp. In addition, negatively regulatory region may exist from -1782 to -1295 bp.[1]References
- Identification and characterization of a glucose-responsiveness region upstream of human insulin gene in transfected HIT-T 15 cells. Ohtani, K., Shimizu, H., Kato, Y., Mori, M. Biochem. Biophys. Res. Commun. (1998) [Pubmed]
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