Cooperation of protein disulfide isomerase and redox environment in the regulation of NF-kappaB and AP1 binding to DNA.
Most transcription factors are multimeric complexes whose subunits depend on strict conformation requirements to form the active unit. Among these requirements is the presence of appropriate sulfhydryl interactions that are critical to transcription factor binding to cognate DNA recognition sites. Our experiments now suggest that modulation of these sulfhydryls may involve the action of thiol-modifying oxido-reductases such as protein disulfide isomerase (PDI). Electrophoretic mobility shift titration experiments incorporating different ratios of GSH:GSSG indicated that changes in GSH and GSSG concentrations corresponding to redox potential differences of as little as +/- 15 mV enabled or abolished binding of NF-kappaB and AP1 to their cognate DNA sites. Moreover, this binding range was modulated significantly by the addition of purified protein disulfide isomerase (PDI). Collectively, these results suggest that a reversible oxidation/reduction signalling pathway may exist in the cell whereby localized changes in redox potentials and/or oxido-reductase activity can be functionally relevant in the regulation of critical gene expression events.
Department of Biology and The Institue for Biomolecular Studies, The Catholic University of America, Washington, D.C. 20064, USA.
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Greene
D033961:Adaptor Protein Complex 1; D033965:Adaptor Protein Complex alpha Subunits; D033942:Adaptor Proteins, Vesicular Transport; D002457:Cell Extracts; D002467:Cell Nucleus / Q000378:metabolism; D004273:DNA, Neoplasm / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology; D004586:Electrophoresis; D005354:Fibrosarcoma; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D019803:Glutathione Disulfide; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007535:Isomerases / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D019704:Protein Disulfide-Isomerases; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000201:enzymology
D033961:Adaptor Protein Complex 1; D033965:Adaptor Protein Complex alpha Subunits; D033942:Adaptor Proteins, Vesicular Transport; D002457:Cell Extracts; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D016328:NF-kappa B; D014157:Transcription Factors; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D005978:Glutathione; D019803:Glutathione Disulfide