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 mu M and 1 mu M, 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 Cys(272) and Cys(154) 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 mu M gold(I) or 10 mu M selenite inhibited expression of an AP-l-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 antiarthiritic action of gold and cancer chemoprevention by selenium.