title: Evidence for 1,25-dihydroxyvitamin D3-independent transactivation by the vitamin D receptor: uncoupling the receptor and ligand in keratinocytes.
abstract: The vitamin D endocrine system plays critical although poorly understood roles in skin. Vitamin D receptor (VDR) knock-out (VDRKO) mice have defects in hair follicle cycling and keratinocyte proliferation leading to epidermal thickening, dermal cyst formation, and alopecia. Surprisingly, skin defects are not apparent in mice lacking 25-hydroxyvitamin D 1alpha-hydroxylase, the enzyme required for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) hormone biosynthesis. These disparate phenotypes indicate that VDR effects in skin are independent of the 1,25(OH)2D3 ligand. However, cellular or molecular data supporting this hypothesis are lacking. Here, we show transcriptional activation of the vitamin D-responsive 24-hydroxylase promoter by VDR in primary keratinocytes that is independent of the 1,25(OH)2D3 ligand. This activity required functional vitamin D-responsive promoter elements as well as an intact VDR DNA binding domain and thus could not be distinguished from 1,25(OH)2D3-dependent VDR transactivation. The 1,25(OH)2D3-independent activation of VDR was also observed in keratinocytes from 1alpha-hydroxylase knock-out mice, indicating that it is not due to endogenous 1,25(OH)2D3 production. Mammalian two-hybrid studies showed strong, 1,25(OH)2D3-independent interaction between VDR and retinoid X receptors in primary keratinocytes, indicating that enhanced heterodimerization of these receptors was involved. Indeed, this 1,25(OH)2D3-independent VDR-RXR heterodimerization was sufficient to drive transactivation by VDR(L233S), an inactive ligand binding mutant of VDR that was previously shown to rescue the skin phenotype of VDR null mice. Cumulatively, these studies support the concept that transactivation by VDR in keratinocytes may be uncoupled from the 1,25(OH)2D3 ligand.
authors.affiliation: Department of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.
authors.forename: Tara I
authors.initials: TI
authors.lastname: Ellison
authors.forename: Richard L
authors.initials: RL
authors.lastname: Eckert
authors.forename: Paul N
authors.initials: PN
authors.lastname: MacDonald
mesh_terms: D000818:Animals; D001665:Binding Sites; D002117:Calcitriol / Q000096:biosynthesis / Q000494:pharmacology*; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D018167:Receptors, Calcitriol / Q000235:genetics / Q000378:metabolism*; D020218:Response Elements; D047488:Retinoid X Receptors / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects* / Q000235:genetics*; D065668:Vitamin D3 24-Hydroxylase
chemical_list: D008024:Ligands; D018167:Receptors, Calcitriol; D047488:Retinoid X Receptors; D004247:DNA; D013250:Steroid Hydroxylases; D065668:Vitamin D3 24-Hydroxylase; D002117:Calcitriol