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J Biol Chem, Vol. 274, Issue 5, 2750-2757, January 29, 1999
From the Glucose-6-phosphate dehydrogenase (G6PD) is the
key enzyme of the pentose phosphate pathway that is responsible for the
generation of NADPH, which is required in many detoxifying reactions.
We have recently demonstrated that G6PD expression is induced by a
variety of chemical agents acting at different steps in the biochemical pathway controlling the intracellular redox status. Although we obtained evidence that the oxidative stress-mediated enhancement of G6PD expression is a general phenomenon, the
functional significance of such G6PD induction after oxidant insult is
still poorly understood. In this report, we used a GSH-depleting drug that determines a marked decrease in the intracellular pool of reduced
glutathione and a gradual but notable increase in G6PD expression. Both
effects are seen soon after drug addition. Once G6PD activity has
reached the maximum, the GSH pool is restored. We suggest and also
provide the first direct evidence that G6PD induction serves to
maintain and regenerate the intracellular GSH pool. We used HeLa cell
clones stably transfected with the human G6PD gene that display higher
G6PD activity than the parent HeLa cells. Although the activities of
glutathione peroxidase, glutathione reductase, and catalase were
comparable in all strains, the concentrations of GSH were significantly
higher in G6PD-overexpressing clones. A direct consequence of GSH
increase in these cells is a decreased reactive oxygen species
production, which makes these cells less sensitive to the oxidative
burst produced by external stimuli. Indeed, all clones that
constitutively overexpress G6PD exhibited strong protection against
oxidants-mediated cell killing. We also observe that NF-
Enhanced Glutathione Levels and Oxidoresistance Mediated by
Increased Glucose-6-phosphate Dehydrogenase Expression
,,,,
International Institute of Genetics and
Biophysics,
B
activation, in response to tumor necrosis factor-
treatment, is
strongly reduced in human HeLa cells overexpressing G6PD.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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