Inhibition of AP-1 DNA binding by nitric oxide involving conserved cysteine residues in Jun and Fos.
Nitric oxide (NO), which has diverse biological effects, can modulate AP-1 activity. Since DNA binding of Jun-Jun and Jun-Fos dimers is regulated in vitro by redox control involving conserved cysteines, we hypothesized that the action of NO is mediated via these residues. We performed electrophoretic mobility-shift analyses using Jun and Fos recombinant proteins and NO solutions. Cysteine-to-serine mutants showed that the inhibition of AP-1 activity following NO treatment was dependent on the presence of Cys7272 and Cys154 in the DNA binding domain of Jun and Fos, respectively. The inhibitory effect of NO was reversed by DTT and the thioredoxin system. Our results demonstrate that NO mediates its inhibitory effect by reacting specifically with the conserved cysteine residues in Jun and Fos.
Nikitovic
D
D
Department of Medical Biochemistry and Biophysics, Karolinska Institute, Sweden.
Holmgren
A
A
Spyrou
G
G
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D003545:Cysteine / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D009569:Nitric Oxide / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011485:Protein Binding / Q000187:drug effects; D011499:Protein Processing, Post-Translational; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism*; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000378:metabolism*
D004268:DNA-Binding Proteins; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D018808:Transcription Factor AP-1; D009569:Nitric Oxide; D013879:Thioredoxins; D004247:DNA; D003545:Cysteine; D004229:Dithiothreitol