Inhibition of AP-1 binding and transcription by gold and selenium involving conserved cysteine residues in Jun and Fos.
Gold(I) salts and selenite, which have diverse therapeutic and biological effects, are noted for their reactivity with thiols. Since the binding of Jun-Jun and Jun-Fos dimers to the AP-1 DNA binding site is regulated in vitro by a redox process involving conserved cysteine residues, we hypothesized that some of the biological actions of gold and selenium are mediated via these residues. In electrophoretic mobility-shift analyses, AP-1 DNA binding was inhibited by gold(I) thiolates and selenite, with 50% inhibition occurring at approximately 5 microM and 1 microM, respectively. Thiomalic acid had no effect in the absence of gold(I), and other metal ions inhibited at higher concentrations, in a rank order correlating with their thiol binding affinities. Cysteine-to-serine mutants demonstrated that these effects of gold(I) and selenite require Cys272 and Cys154 in the DNA-binding domains of Jun and Fos, respectively. Gold(I) thiolates and selenite did not inhibit nonspecific protein binding to the AP-1 site and were at least an order of magnitude less potent as inhibitors of sequence-specific binding to the AP-2, TFIID, or NF1 sites compared with the AP-1 site. In addition, 10 microM gold(I) or 10 microM selenite inhibited expression of an AP-1-dependent reporter gene, but not an AP-2-dependent reporter gene. These data suggest a mechanism regulating transcription factor activity by inorganic ions which may contribute to the known antiarthritic action of gold and cancer chemoprevention by selenium.
Handel
M L
ML
Cancer Biology Division, Garvan Institute of Medical Research, Sydney, New South Wales, Australia.
Watts
C K
CK
deFazio
A
A
Day
R O
RO
Sutherland
R L
RL
D000595:Amino Acid Sequence; D001665:Binding Sites; D001943:Breast Neoplasms; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000378:metabolism; D017124:Conserved Sequence; D003545:Cysteine; D004305:Dose-Response Relationship, Drug; D006046:Gold / Q000494:pharmacology*; D006052:Gold Sodium Thiomalate / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D016760:Proto-Oncogene Proteins c-fos / Q000737:chemistry / Q000378:metabolism*; D016755:Proto-Oncogene Proteins c-jun / Q000737:chemistry / Q000378:metabolism*; D018038:Sodium Selenite / Q000494:pharmacology*; D018808:Transcription Factor AP-1 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D006052:Gold Sodium Thiomalate; D006046:Gold; D015500:Chloramphenicol O-Acetyltransferase; D018038:Sodium Selenite; D003545:Cysteine