H2O2 and antioxidants have opposite effects on activation of NF-kappa B and AP-1 in intact cells: AP-1 as secondary antioxidant-responsive factor

EMBO J. 1993 May;12(5):2005-15.

Abstract

We show that AP-1 is an antioxidant-responsive transcription factor. DNA binding and transactivation by AP-1 were induced in HeLa cells upon treatment with the antioxidants pyrrolidine dithiocarbamate (PDTC) and N-acetyl-L-cysteine (NAC), and upon transient expression of the antioxidative enzyme thioredoxin. While PDTC and NAC enhanced DNA binding and transactivation of AP-1 in response to phorbol ester, the oxidant H2O2 suppressed phorbol ester activation of the factor. H2O2 on its own was only a weak inducer of AP-1. Activation of AP-1 by PDTC was dependent on protein synthesis and involved transcriptional induction of c-jun and c-fos genes. Transcriptional activation of c-fos by PDTC was conferred by the serum response element, suggesting that serum response factor and associated proteins function as primary antioxidant-responsive transcription factors. In the same cell line, the oxidative stress-responsive transcription factor NF-kappa B behaved in a manner strikingly opposite to AP-1. DNA binding and transactivation by NF-kappa B were strongly activated by H2O2, while the antioxidants alone were ineffective. H2O2 potentiated the activation of NF-kappa B by phorbol ester, while PDTC and NAC suppressed PMA activation of the factor. PDTC did not influence protein kinase C (PKC) activity and PKC activation by PMA, indicating that the antioxidant acted downstream of and independently from PKC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / pharmacology*
  • Base Sequence
  • DNA
  • Genes, fos
  • Genes, jun
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / pharmacology*
  • Molecular Sequence Data
  • NF-kappa B / metabolism*
  • Oligodeoxyribonucleotides
  • Proto-Oncogene Proteins c-jun / metabolism*
  • Pyrrolidines / pharmacology
  • Regulatory Sequences, Nucleic Acid
  • Tetradecanoylphorbol Acetate / pharmacology
  • Thiocarbamates / pharmacology
  • Thioredoxins / biosynthesis
  • Transcription, Genetic
  • Transcriptional Activation

Substances

  • Antioxidants
  • NF-kappa B
  • Oligodeoxyribonucleotides
  • Proto-Oncogene Proteins c-jun
  • Pyrrolidines
  • Thiocarbamates
  • pyrrolidine dithiocarbamic acid
  • Thioredoxins
  • DNA
  • Hydrogen Peroxide
  • Tetradecanoylphorbol Acetate