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Diabetes 52:403-408, 2003
© 2003 by the American Diabetes Association, Inc.

BETA2 Activates Transcription From the Upstream Glucokinase Gene Promoter in Islet ß-Cells and Gut Endocrine Cells

J. Michael Moates1,2,3, Sarmistha Nanda1, Michelle A. Cissell4, Ming-Jer Tsai1,2, and Roland Stein4

1 Department of Medicine, Baylor College of Medicine, Houston, Texas
2 Department of Molecular & Cellular Biology, Baylor College of Medicine, Houston, Texas
3 Department of Veterans Affairs, Houston, Texas
4 Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, Tennessee

Glucokinase (GK) gene transcription initiates in the islet (ß-cell), gut, and brain from promoter sequences residing ~35 kbp upstream from those used in liver. Expression of ßGK is controlled in ß-cells by cell-enriched (i.e. pancreatic duodenal homeobox 1 [PDX-1]) and ubiquitously (i.e., Pal) distributed factors that bind to and activate from conserved sequence motifs within the upstream promoter region (termed ßGK). Here, we show that a conserved E-box element also contributes to control in the islet and gut. ßGK promoter-driven reporter gene activity was diminished by mutating the specific sequences involved in E-box-mediated basic helix-loop-helix factor activator binding in islet ß-cells and enteroendocrine cells. Gel shift assays demonstrated that the ßGK and insulin gene E-box elements formed the same cell-enriched (BETA2:E47) and generally distributed (upstream stimulatory factor [USF]) protein-DNA complexes. ßGK E-box-driven activity was stimulated in cotransfection assays performed in baby hamster kidney (BHK) cells with BETA2 and E47, but not USF. Chromatin immunoprecipitation assays performed with BETA2 antisera showed that BETA2 occupies the upstream promoter region of the endogenous ßGK gene in ß-cells. We propose that BETA2 (also termed NeuroD1) regulates ßGK promoter activity.


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