Journal
AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY
Volume 278, Issue 4, Pages G591-G603Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajpgi.2000.278.4.G591
Keywords
intestine; transcription; glucose-galactose malabsorption; Sp2; Sp3
Categories
Funding
- NICHD NIH HHS [HD-34706] Funding Source: Medline
- NIDDK NIH HHS [DK-44582] Funding Source: Medline
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The Na+-glucose cotransporter (SGLT1) is expressed primarily by small intestinal epithelial cells and transports the monosaccharides glucose and galactose across the apical membrane. Here we describe the isolation and characterization of 5.3 kb of the 5'-flanking region of the SGLT1 gene by transiently transfecting reporter constructs into a variety of epithelial cell lines. A fragment (nt -235 to +22) of the promoter showed strong activity in the intestinal cell line Caco-2 but was inactive in a nonintestinal epithelial cell line (Chinese hamster ovary). Within this region, three cis-elements, a hepatocyte nuclear factor-1 (HNF-1) and two GC box sites are critical for maintaining the gene's basal level of expression. The two GC boxes bind to several members of the Spl family of transcription factors and, in the presence of HNF-1, synergistically upregulate transactivation of the promoter. A novel 16-bp element just downstream of one GC box was also shown to influence the interaction of Spl to its binding site. In summary, we report the identification and characterization of the human SGLT1 minimal promoter and the critical role that HNF-1 and Spl-multigene members have in enhancing the basal level of its transcription in Caco-2 cells.
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