Redox regulation of c-Jun DNA binding by reversible S-glutathiolation.
Redox control of the transcription factor c-Jun maps to a single cysteine in its DNA binding domain. However, the nature of the oxidized state of this cysteine and, thus, the potential molecular mechanisms accounting for the redox regulation of c-Jun DNA binding remain unclear. To address this issue, we have analyzed the purified recombinant c-Jun DNA binding domain for redox-dependent thiol modifications and concomitant changes in DNA binding activity. We show that changes in the ratio of reduced to oxidized glutathione provide the potential to oxidize c-Jun sulfhydryls by mechanisms that include both protein disulfide formation and S-glutathiolation. We provide evidence that S-glutathiolation, which is specifically targeted to the cysteine residue located in the DNA binding site of the protein, may account for the reversible redox regulation of c-Jun DNA binding. Furthermore, based on a molecular model of the S-glutathiolated protein, we discuss the structural elements facilitating S-glutathiolation and how this modification interferes with DNA binding. Given the structural similarities between the positively charged cysteine-containing DNA binding motif of c-Jun and the DNA binding site of related oxidant-sensitive transcriptional activators, the unprecedented phenomenon of redox-triggered S-thiolation of a transcription factor described in this report suggests a novel role for protein thiolation in the redox control of transcription.
Klatt
P
P
Departamento de Estructura y Función de Proteínas, Instituto Reina Sofía de Investigaciones Nefrológicas, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28006 Madrid, Spain. pklatt@cib.csic.es
Molina
E P
EP
De Lacoba
M G
MG
Padilla
C A
CA
Martinez-Galesteo
E
E
Barcena
J A
JA
Lamas
S
S
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001483:Base Sequence; D001665:Binding Sites; D003196:Computer Graphics; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D006706:Homeostasis; D006801:Humans; D007700:Kinetics; D016350:Leucine Zippers; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D016755:Proto-Oncogene Proteins c-jun / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D016755:Proto-Oncogene Proteins c-jun; D011994:Recombinant Proteins; D004247:DNA; D005978:Glutathione