Transcriptional regulation of the ovine intestinal Na+/glucose cotransporter SGLT1 gene. Role of HNF-1 in glucose activation of promoter function.

Abstract

Dietary sugars D-glucose and D-galactose are transported across the intestinal brush-border membrane by the Na+/glucose cotransporter, SGLT1. In various species studied, it has been shown that the activity, and expression, of intestinal SGLT1 is regulated by dietary sugars. We report in this paper that regulation of the intestinal SGLT1 gene by lumenal sugar is due, in part, to an increase in transcription. Using deletion analyses of the -66/+21-bp fragment, we have identified the minimal region of the ovine SGLT1 promoter able to support transcription. Site-directed mutagenesis of the hepatic nuclear factor-1 (HNF-1) consensus motif within this domain eliminates basal promoter function. In addition, we show direct evidence for glucose-induced activation of the -66/+21-bp promoter region. There is a co-ordinated decline in the abundance of ovine intestinal HNF-1 and SGLT1 transcripts during transition from preruminant to adult ruminant. This decline is recovered after glucose infusion of adult sheep intestine. Similarly, as shown using DNA mobility-shift assays, the intensity of the HNF-1-binding complex to the target promoter sequence decreases during maturation of the animal; this is restored after intestinal sugar infusion. These data indicate that HNF-1 plays an important role in the glucose responsiveness of the ovine SGLT1 gene. This is the first report of in vitro glucose-induced activation of the intestinal SGLT1 promoter and identification of a glucose-responsive region of the ovine SGLT1 promoter.