Cloning and characterization of the mouse glucokinase gene locus and identification of distal liver-specific DNase I hypersensitive sites

Journal Article · Tue Oct 10 04:00:00 EDT 1995 · Genomics

DOI:https://doi.org/10.1006/geno.1995.9943· OSTI ID:443874

Postic, C; Niswender, K D; Shelton, K D; Pettepher, C C; Granner, D K; Magnuson, M A ^[1]

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1. Vanderbilt Univ., Nashville, TN (United States); and others

We cloned and characterized an 83-kb fragment of mouse genomic DNA containing the entire glucokinase (GK) gene. The 11 exons of the gene span a total distance of 49 kb, with exons 1{beta} and 1L being separated by 35 kb. A total of 25,266 bp of DNA sequence information was determined: from {approximately}-9.2 to {approximately}+15 kb (24,195 bp), relative to the hepatocyte transcription start site, and from -335 to -736 bp (1071 bp), relative to the transcription start site in {beta} cells. These sequences revealed that mouse GK is >94% identical to rat and human GK. Mouse hepatic GK mRNA is regulated by fasting and refeeding, as also occurs in the rat. Alignment of the upstream and downstream promoter regions of the mouse, rat, and human genes revealed several evolutionarily conserved regions that may contribute to transcriptional regulation. However, fusion gene studies in transgenic mice indicate that the conserved regions near the transcription start site in hepatocytes are themselves not sufficient for position-independent expression in liver. Analysis of the chromatin structure of a 48-kb region of the mouse gene using DNase I revealed eight liver-specific hypersensitive sites whose locations ranged from 0.1 to 36 kb upstream of the liver transcription start site. The availability of a single, contiguous DNA fragment containing the entire mouse GK gene should allow further studies of cell-specific expression of GK to be performed. 46 refs., 8 figs.

• Sponsoring Organization: USDOE
• OSTI ID: 443874
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 3 Vol. 29; ISSN GNMCEP; ISSN 0888-7543
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
AMINO ACID SEQUENCE
BIOLOGICAL EVOLUTION
DNA SEQUENCING
DNA-ASE
DNA-CLONING
GENE REGULATION
GENE REPRESSORS
GENETIC MAPPING
HEXOKINASE
HUMAN POPULATIONS
LIVER CELLS
MICE
POLYMERASE CHAIN REACTION
RATS
STRUCTURE-ACTIVITY RELATIONSHIPS
TRANSCRIPTION