Thioredoxin regulates the DNA binding activity of NF-kappa B by reduction of a disulphide bond involving cysteine 62.
A role for redox regulation in activation of the NF-kappa B transcription factor was suggested by the observation that DNA binding activity of free protein, but not preformed DNA-protein complex, is inhibited by -SH modifying agents but enhanced by reducing agents. Mutagenesis of conserved cysteine residues in the p50 subunit identified amino acid 62 as being important for DNA binding, as a serine substitution at this position reduces DNA binding affinity, but renders the protein insensitive to -SH modifying agents. DNA binding activity of the wild type protein but not the amino acid 62 mutant was also stimulated by thioredoxin while detection of disulphide cross linked dimers in p50 but not the amino acid 62 mutant suggests that thioredoxin stimulates DNA binding by reduction of a disulphide bond involving cysteine 62. The physiological relevance of these findings was supported by the observation that cotransfection of a plasmid expressing human thioredoxin and an HIV LTR driven reporter construct resulted in an NF-kappa B dependent increase in expression of the reporter gene. Thus modification of p50 by thioredoxin, a gene induced by stimulation of T-lymphocytes in parallel with NF-kappa B translocation, is a likely step in the cascade of events leading to full NF-kappa B activation.
Matthews
J R
JR
Division of Biochemistry and Molecular Biology, School of Biological and Medical Sciences, University of St Andrews, Fife, UK.
Wakasugi
N
N
Virelizier
J L
JL
Yodoi
J
J
Hay
R T
RT
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D013601:T-Lymphocytes; D013879:Thioredoxins / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics
D004220:Disulfides; D016328:NF-kappa B; D013879:Thioredoxins; D004247:DNA; D003545:Cysteine