A tyrosine kinase-dependent pathway regulates ligand-dependent activation of the dioxin receptor in human keratinocytes.
Signal transduction by dioxin is mediated by the intracellular basic helix-loop-helix dioxin receptor which, in its ligand-activated state, binds to target DNA as a heteromeric complex with the partner factor Arnt. In contrast, the repressed form of the receptor is a complex with hsp90 which appears to maintain the receptor in an inducible conformation. In human keratinocytes dioxin receptor activation has previously been shown to depend on phosphorylation processes. To further dissect mechanisms regulating dioxin receptor function the importance of tyrosine phosphorylation was investigated by the use of specific tyrosine kinase inhibitors. Here we report that the inhibitor genistein inhibited dioxin-dependent induction of expression of the target gene cytochrome P-450IA1. This effect was rapid and reversible and did not lead to altered levels of dioxin receptor protein. Analyses of dioxin receptor or Arnt fusion proteins that function independently of one another showed that the target for genistein action was the dioxin receptor, and, more specifically, a region of the receptor harboring its ligand-binding domain. In addition, function of an unrelated transactivator, the glucocorticoid receptor, was inhibited by genistein while a truncated form lacking the ligand-binding domain was not. A common denominator between the ligand-binding domains of both receptors is their ability to interact with hsp90. Importantly, co-immunoprecipitation experiments showed that genistein inhibited ligand-induced release of hsp90 from the glucocorticoid receptor. Thus, the interaction of these transactivators with hsp90 may be regulated by a tyrosine kinase-dependent pathway.
Gradin
K
K
Center for Nutrition and Toxicology, Karolinska Institute, Novum, Huddinge, Sweden.
Whitelaw
M L
ML
Toftgård
R
R
Poellinger
L
L
Berghard
A
A
D000818:Animals; D001483:Base Sequence; D016227:Benzoquinones; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects; D019833:Genistein; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology; D015603:Keratinocytes / Q000378:metabolism*; D047029:Lactams, Macrocyclic; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011809:Quinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017828:Rifabutin / Q000031:analogs & derivatives
D016227:Benzoquinones; D017931:DNA Primers; D006360:Heat-Shock Proteins; D007529:Isoflavones; D047029:Lactams, Macrocyclic; D008024:Ligands; D011809:Quinones; D012333:RNA, Messenger; D018336:Receptors, Aryl Hydrocarbon; D011965:Receptors, Glucocorticoid; D011993:Recombinant Fusion Proteins; D017828:Rifabutin; C004742:daidzein; C020754:herbimycin; D003577:Cytochrome P-450 Enzyme System; D019833:Genistein; D011505:Protein-Tyrosine Kinases