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Volume 272, Number 52, Issue of December 26, 1997 pp. 33367-33372

Transcriptional Activation of the Glut1 Gene in Response to Oxidative Stress in L6 Myotubes

(Received for publication, June 11, 1997, and in revised form, October 8, 1997)

Nitzan Kozlovsky , Assaf Rudich , Ruth Potashnik , Yousuke Ebina ^§ , Takashi Murakami ^§ and Nava Bashan

From the  Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-sheva 84105, Israel and the ^§ Department of Enzyme Genetics, Institute for Enzyme Research, University of Tokushima, Kuramoto-cho 770, Japan

Exposure of L6 myotubes to prolonged low grade oxidative stress results in increased Glut1 expression at both the protein and mRNA levels, leading to elevated glucose transport activity. To further understand the cellular mechanisms responsible for this adaptive response, the Glut1 transcription rate and mRNA stability were assessed. Nuclear run-on assays revealed 2.0- and 2.4-fold increases in Glut1 transcription rates in glucose oxidase- and xanthine/xanthine oxidase-pretreated cells, respectively. Glut1 mRNA stability was increased with both treatments compared with the control (t1/2 = 7.8 ± 1.3, 6.0 ± 2.0, and 2.4 ± 0.5 h, respectively). The serum-responsive element and AP-1 (but not the cAMP-responsive element) showed increased binding capacity following oxidative stress. Both activation of AP-1 binding and elevation of Glut1 mRNA were prevented by cycloheximide. The involvement of enhancer 1 of the Glut1 gene was demonstrated using transfected 293 cells. Induction of Glut1 mRNA in response to oxidative stress differed from its activation by chronic insulin exposure as demonstrated by the ability of rapamycin to inhibit the latter without an effect on the former. In conclusion, oxidative stress increases the Glut1 transcription rate by mechanisms that may involve activation of AP-1 binding to enhancer 1 of the Glut1 gene.

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