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Diabetologia

, Volume 38, Issue 1, pp 112–115 | Cite as

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

  • B. Leibiger
  • I. B. Leibiger
Rapid Communication

Summary

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 βTC-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 βTC-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 delineatecis-elements within the region −220/+1. Five specifically protected areas could be defined. [Diabetologia (1995) 38: 112–115]

Key words

Gene expression regulation insulinoma glucose transporter transcription promoter 

Abbreviations

GLUT 2

Glucose transporter

PCR

polymearase chain reaction

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References

  1. 1.
    Matschinsky FM (1990) Glucokinase as glucose sensor and metabolic signal generator in pancreatic β-cells and hepatocytes. Diabetes 39: 647–652PubMedGoogle Scholar
  2. 2.
    Malaisse WJ (1994) Is glucokinase the beta-cell glucoreceptor? Diabetologia 37: 442.CrossRefPubMedGoogle Scholar
  3. 3.
    Randle PJ (1993) Glucokinase and candidate genes for type 2 (non-insulin-dependent) diabetes mellitus. Diabetologia 36: 269–275CrossRefPubMedGoogle Scholar
  4. 4.
    Thorens B, Sarkar HK, Kaback HR, Lodish JF (1988) Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney and β-pancreatic islet cells. Cell 55: 281–290CrossRefPubMedGoogle Scholar
  5. 5.
    Mueckler M (1994) Facilitative glucose transporters. FEBS Lett 219: 713–725Google Scholar
  6. 6.
    MacDonald MJ, McKenize DI, Walker TM, Kaysen JH (1992) Lack of gluconeogenesis in pancreatic islets: expression of gluconeogenic enzyme genes in islets. Horm Metab Res 24: 158–160PubMedGoogle Scholar
  7. 7.
    Takeda J, Kayano T, Fukomoto H, Bell GI (1993) Organization of the human GLUT 2 (pancreatic β-cell and hepatocyte) glucose transporter gene. Diabetes 42: 773–777PubMedGoogle Scholar
  8. 8.
    Boam DSW, Clark AR, Docherty K (1990) Positive and negative regulation of the human insulin gene by multiple trans-acting factors. J Biol Chem 265: 8285–8296PubMedGoogle Scholar
  9. 9.
    Leibiger IB, Walther R, Pett U, Leibiger B (1994) Positive and negative regulatory elements are involved in transcriptional control of the rat glucokinase gene in the insulin-producing cell line HIT M2.2.2. FEBS Lett 337: 161–166CrossRefPubMedGoogle Scholar
  10. 10.
    Ohlsson H, Edlund T (1986) Sequence-specific interactions of nuclear factors with the insulin gene enhancer. Cell 45: 35–44CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • B. Leibiger
  • I. B. Leibiger
    • 1
  1. 1.Institute of Biochemistry, School of MedicineUniversity of GreifswaldGreifswaldGermany

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