title: Metal mediated sterol receptor-DNA complex association and dissociation determined by electrospray ionization mass spectrometry.
abstract: The vitamin D receptor (VDR) binds to specific DNA sequences termed vitamin D response elements (VDREs) thereby enhancing or repressing transcription. We have used electrospray ionization mass spectrometry to examine the interaction between the DNA-binding domain of the vitamin D receptor (VDR DBD) with a double-stranded DNA (dsDNA) sequence containing the VDRE from the mouse osteopontin gene. The VDR DBD was shown to bind to the appropriate DNA sequence only when bound to 2 moles of zinc (Zn2+) or cadmium (Cd2+) per mole of protein. Additional binding of Zn2+ or Cd2+ by the protein caused the protein to dissociate from the dsDNA. These results show that the VDR DBD/DNA metal-dependent association occurs when the receptor is occupied by 2 moles of Zn2+ per mole of protein and that further binding of Zn2+ to the protein causes dissociation of the complex.
authors.affiliation: Nephrology Research Unit, Mayo Clinic/Foundation, Rochester, MN 55905, USA.
authors.forename: T D
authors.initials: TD
authors.lastname: Veenstra
authors.forename: L M
authors.initials: LM
authors.lastname: Benson
authors.forename: T A
authors.initials: TA
authors.lastname: Craig
authors.forename: A J
authors.initials: AJ
authors.lastname: Tomlinson
authors.forename: R
authors.initials: R
authors.lastname: Kumar
authors.forename: S
authors.initials: S
authors.lastname: Naylor
mesh_terms: D000818:Animals; D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D053495:Osteopontin; D018167:Receptors, Calcitriol / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D012795:Sialoglycoproteins / Q000235:genetics / Q000378:metabolism; D015032:Zinc / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D018167:Receptors, Calcitriol; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D002104:Cadmium; D053495:Osteopontin; D004247:DNA; D015032:Zinc