Nrf2-dependent activation of the antioxidant responsive element by tert-butylhydroquinone is independent of oxidative stress in IMR-32 human neuroblastoma cells.
The antioxidant responsive element (ARE) is a cis-acting regulatory element located in the 5'-flanking region of several genes encoding phase II detoxification enzymes, including NAD(P)H:quinone oxidoreductase (NQO1). We report here that activation of the NQO1 ARE by tert-butylhydroquinone (tBHQ) is dependent on Nrf2 and not oxidative stress in IMR-32 human neuroblastoma cells. Overexpression of wild-type Nrf2 activated ARE in a dose-dependent manner, and ARE activation by tBHQ or diethyl maleate (DEM) was inhibited by dominant/negative Nrf2 not by dominant/negative c-Jun. According to our observation, the palindromic sequence (5' to the core) and the GC box in the ARE core sequence are essential for maximal inducibility by tBHQ or DEM. Overexpression of Nrf2 selectively activated wild-type ARE up to 24 h. In addition, a dramatic nuclear translocation of Nrf2 by tBHQ supports a role for Nrf2 in ARE activation. Although oxidative stress is hypothesized to be a major driving force for ARE activation, pretreatment of antioxidant or antioxidant enzyme did not block tBHQ-mediated ARE activation. In contrast, ARE activation by DEM was inhibited by antioxidants or catalase. These results suggest that ARE activation signals from tBHQ and DEM converge at Nrf2 transcription factor through independent mechanisms.
Lee
J M
JM
School of Pharmacy, University of Wisconsin, 425 N. Charter St., Madison, WI 53706, USA.
Moehlenkamp
J D
JD
Hanson
J M
JM
Johnson
J A
JA
D000975:Antioxidants / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D016758:Genes, jun; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology*; D007700:Kinetics; D016350:Leucine Zippers; D008298:Maleates / Q000494:pharmacology; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000235:genetics*; D051267:NF-E2-Related Factor 2; D009447:Neuroblastoma; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D000975:Antioxidants; D004268:DNA-Binding Proteins; D006873:Hydroquinones; D008298:Maleates; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; C018855:2-tert-butylhydroquinone; D016660:NAD(P)H Dehydrogenase (Quinone); C014476:diethyl maleate