Isolation and characterization of a genomic region upstream from the ovine Na+/d-glucose cotransporter (SGLT1) cDNA.
d-Glucose and non-metabolisable analogues of D-glucose regulate the expression of intestinal SGLT1 at both transcriptional and post-transcriptional levels. In order to investigate the molecular mechanisms involved in the transcriptional regulation of the ovine intestinal SGLT1 gene, we have isolated an upstream element of about 1 kb in size. This DNA fragment contains a TATA box motif, 48 bp upstream of the transcriptional start site and includes transcription factor binding sites for HNF-1 and AP-2. We have shown that the ovine SGLT1 promoter fragment can drive the transcription of a reporter gene when transfected into the epithelial cell lines STC-1 and LLC-PK1, which endogenously express SGLT1. Deletion analyses of the promoter indicate that -66/+21 bp proximal sequence directs the highest level of reporter gene activity. There are one and possibly two sites of transcriptional suppression.
Wood
I S
IS
Department of Veterinary Preclinical Sciences, University of Liverpool, Brownlow Hill, L69 3BX, United Kingdom.
Allison
G G
GG
Shirazi-Beechey
S P
SP
D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism; D004847:Epithelial Cells; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics; D006041:Goats / Q000235:genetics*; D051537:Hepatocyte Nuclear Factor 1; D051538:Hepatocyte Nuclear Factor 1-alpha; D051539:Hepatocyte Nuclear Factor 1-beta; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000235:genetics*; D009687:Nuclear Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012097:Repressor Proteins / Q000502:physiology; D020218:Response Elements / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D051269:Sodium-Glucose Transporter 1; D016385:TATA Box / Q000235:genetics; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
D020121:5' Untranslated Regions; D004268:DNA-Binding Proteins; C496487:HNF1A protein, human; C496495:HNF1B protein, human; D051538:Hepatocyte Nuclear Factor 1-alpha; D008562:Membrane Glycoproteins; D009004:Monosaccharide Transport Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; C495674:SLC5A1 protein, human; D051269:Sodium-Glucose Transporter 1; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D051537:Hepatocyte Nuclear Factor 1; D051539:Hepatocyte Nuclear Factor 1-beta