Metal mediated sterol receptor-DNA complex association and dissociation determined by electrospray ionization mass spectrometry

• Published in: Nature biotechnology 1998-03, Vol.16 (3), p.262-266
• Main Authors: Veenstra, Timothy D, Benson, Linda M, Craig, Theodore A, Tomlinson, Andy J, Kumar, Rajiv, Naylor, Stephen
• Format: Article
• Language: English
• Subjects: Agriculture; Animals; Binding Sites; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Cadmium - metabolism; Cadmium - pharmacology; DNA - chemistry; DNA - metabolism; Fundamental and applied biological sciences. Psychology; Interactions. Associations; Intermolecular phenomena; Life Sciences; Mass Spectrometry - methods; Mice; Molecular biophysics; Osteopontin; Receptors, Calcitriol - chemistry; Receptors, Calcitriol - drug effects; Receptors, Calcitriol - metabolism; research-article; Sialoglycoproteins - genetics; Sialoglycoproteins - metabolism; Zinc - metabolism; Zinc - pharmacology
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt0398-262

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 DMA-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 (Zn 2+ ) or cadmium (Cd 2+ ) per mole of protein. Additional binding of Zn 2+ or Cd 2+ 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 Zn 2+ per mole of protein and that further binding of Zn 2+ to the protein causes dissociation of the complex.