Open AccessA nuclear protein essential for binding of rat 1,25-dihydroxyvitamin D3 receptor to its response elements.
Author(s) -
Troy K. Ross,
Valerie Moss,
Jean M. Prahl,
Hector F. DeLuca
Publication year - 1992
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.89.1.256
Subject(s) - recombinant dna , nuclear receptor , calcitriol receptor , biology , receptor , electrophoretic mobility shift assay , microbiology and biotechnology , vitamin d binding protein , binding protein , skeletal muscle , nuclear protein , transcription factor , vitamin , biochemistry , endocrinology , gene
Recombinant 1,25-dihydroxyvitamin D3 receptor from a baculovirus expression system requires a mammalian-derived nuclear accessory protein for binding to a vitamin D response element (DRE). This was established by electrophoretic mobility shift analyses using radiolabeled DNA probes consisting of DREs from two vitamin D-responsive genes. Mammalian nuclear extract was also required for the binding of wild-type porcine vitamin D receptor to a DRE. Surprisingly, the accessory factor-dependent formation of receptor-DRE complex was independent of exogenous 1,25-dihydroxyvitamin D3. A 59- to 64-kDa accessory protein from porcine intestinal nuclear extract was identified by size-exclusion chromatography. Nuclear extracts from rat liver and kidney contained accessory factor, whereas smaller amounts were detected in heart muscle. Spleen and skeletal muscle contained no detectable accessory factor.