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J Biol Chem, Vol. 274, Issue 12, 8181-8190, March 19, 1999

Identification of a Glucose Response Element in the Promoter of the Rat Glucagon Receptor Gene

Laurence Portois, Barbara Maget, Michèle Tastenoy, Jason Perret, and Michal Svoboda

From the Department of Biochemistry and Nutrition, Medical School, Université Libre de Bruxelles, Building G/E, CP 611, 808 Route de Lennik, B-1070 Brussels, Belgium

We cloned the 5' upstream region of the rat glucagon receptor gene, demonstrating that the 5' noncoding domain of the glucagon receptor mRNA contained two untranslated exons of 131 and 166 nucleotides (nt), respectively, separated by two introns of 0.6 and 3.2 kilobase pairs. We also observed an alternative splicing involving the 166-base pair exon. Cloning of up to 2 kilobase pairs of the newly identified genomic domain and transfection of various constructs driving a reporter gene, in pancreatic islet cell line INS-1, uncovered a strong glucose regulation of the promoter activity of plasmids containing up to nucleotide -868, or more, upstream from the transcriptional start point. This promoter activity displayed threshold-like behavior, with low activity of the promoter below 5 mM glucose, and maximal activation as of 10 mM glucose. This glucose regulation was mapped to a highly palindromic 19-nucleotide region between nt -545 and -527. Indeed, deletion or mutation of this sequence abolished the glucose regulation. This domain contained two palindromic "E-boxes" CACGTG and CAGCTG separated by 3 nt, a feature similar to the "L4 box" found in the pyruvate kinase L gene promoter. This is the first description of a G protein-coupled receptor gene promoter regulated by glucose.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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