Functional analysis of DNA-elements involved in transcriptional control of the human glucose transporter 2 (GLUT 2) gene in the insulin-producing cell line beta TC-3.

The uptake of glucose into pancreatic beta cells as a 'non-rate-limiting-step' is guaranteed by the expression and action of the high-Km glucose transporter 2 (GLUT 2). This transporter is not saturated by physiological plasma glucose levels and hence functions as a "glucose sensor/glucoreceptor". Here we describe DNA-elements of the human GLUT 2 gene promoter which contribute to transcriptional control in the insulin-producing cell line beta TC-3. Nested 5'-as well as 3'-deletions of a DNA-fragment containing up to 1245 bp of the 5'-flanking region and up to 308 bp of the first exon of the human GLUT 2 gene were investigated for their ability to control the expression of a CAT reporter gene in beta TC-3 cells. For tissue-specific transcriptional control 5'-deletional analysis revealed that the region -220/+309 was sufficient. Truncation from the 3'-end from nucleotide +308 to +204 led to a threefold drop in CAT expression. In vitro DNase I footprinting analysis was performed to delineate cis-elements within the region -220/+1. Five specifically protected areas could be defined.