Modulation of transcription factor NF-kappa B binding activity by oxidation-reduction in vitro.
NF-kappa B is a widely used regulator of inducible and tissue-specific gene control. In the cytosol, when complexed to an inhibitory molecule, I kappa B, NF-kappa B is in an inactive form and cannot bind DNA. Activation of cells with appropriate stimuli results in the dissociation of NF-kappa B from I kappa B and its translocation to the nucleus as an active binding protein. We now demonstrate that NF-kappa B binding in vitro can be inhibited by agents that modify free sulfhydryls. Binding is eliminated after treatment with N-ethylmaleimide, an alkylating agent, and diamide, an oxidizing agent. The diamide effect can be reversed by 2-mercaptoethanol. Further, 2-mercaptoethanol acts synergistically with deoxycholate plus Nonidet P-40 in converting inactive cytosolic NF-kappa B to an active DNA-binding form. It is therefore possible that modulation of the redox state of NF-kappa B could represent a post-translational control mechanism for this factor.
Toledano
M B
MB
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesa, MD 20892.
Leonard
W J
WJ
D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D003840:Deoxycholic Acid / Q000494:pharmacology; D003958:Diamide / Q000494:pharmacology; D004357:Drug Synergism; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D008623:Mercaptoethanol / Q000494:pharmacology; D016328:NF-kappa B / Q000737:chemistry / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D017830:Octoxynol; D010084:Oxidation-Reduction; D011092:Polyethylene Glycols / Q000494:pharmacology; D016755:Proto-Oncogene Proteins c-jun; D026362:Serum Response Factor; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D016328:NF-kappa B; D009687:Nuclear Proteins; D016755:Proto-Oncogene Proteins c-jun; D026362:Serum Response Factor; D013438:Sulfhydryl Compounds; D014157:Transcription Factors; D003840:Deoxycholic Acid; D003958:Diamide; D011092:Polyethylene Glycols; D008623:Mercaptoethanol; D017830:Octoxynol; D004247:DNA; C010615:Nonidet P-40; D005033:Ethylmaleimide