Translational control of nrf2 protein in activation of antioxidant response by oxidants.
Nf-E2 related factor-2 (Nrf2) is a basic leucine zipper transcription factor that binds and activates the antioxidant response element (ARE) in the promoters of many antioxidant and detoxification genes. We found that H(2)O(2) treatment caused a rapid increase in endogenous Nrf2 protein level in rat cardiomyocytes. Semiquantitative or real-time reverse transcription-polymerase chain reaction failed to show an increase of Nrf2 mRNA level by H(2)O(2) treatment. Measurements of Nrf2 protein stability excluded the possibility of Nrf2 protein stabilization. Although inhibiting protein synthesis with cycloheximide prevented H(2)O(2) from elevating Nrf2 protein level, RNA synthesis inhibition with actinomycin D failed to do so. Measurements of new protein synthesis with [(35)S]methionine incorporation confirmed that H(2)O(2) increased the translation of Nrf2 protein. Inhibitors of phosphoinositide 3-kinase were able to abolish the induction of Nrf2 protein by H(2)O(2). Although H(2)O(2) increased phosphorylation of p70 S6 kinase, rapamycin failed to inhibit H(2)O(2) from elevating Nrf2 protein. H(2)O(2) also induced phosphorylation of eukaryotic translation initiation factor (eIF) 4E and eIF2alpha within 30 and 10 min, respectively. Inhibiting eIF4E with small interfering siRNA or increasing eIF2alpha phosphorylation with salubrinal did not affect Nrf2 elevation by H(2)O(2). Our data present a novel phenomenon of quick onset of the antioxidant/detoxification response via increased translation of Nrf2 by oxidants. The mechanism underlying such stress-induced de novo protein translation may involve multiple components of translational machinery.
Purdom-Dickinson
Sally E
SE
Department of Pharmacology, University of Arizona College of Medicine, 1501 N. Campbell Ave., Tucson, AZ 85724, USA.
Sheveleva
Elena V
EV
Sun
Haipeng
H
Chen
Qin M
QM
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002478:Cells, Cultured; D003609:Dactinomycin / Q000494:pharmacology; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D039561:Eukaryotic Initiation Factor-4E / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics* / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology*; D010766:Phosphorylation; D014176:Protein Biosynthesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology
D000975:Antioxidants; D015852:Eukaryotic Initiation Factor-2; D039561:Eukaryotic Initiation Factor-4E; D051267:NF-E2-Related Factor 2; D016877:Oxidants; D012333:RNA, Messenger; D003609:Dactinomycin; D008715:Methionine; D006861:Hydrogen Peroxide; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D020123:Sirolimus