Vitamin D receptor interaction with specific DNA requires a nuclear protein and 1,25-dihydroxyvitamin D3.
The regulation of osteocalcin gene expression by 1,25-dihydroxyvitamin D3 is mediated by the vitamin D receptor and a cis-acting DNA response element that has been identified within the 5' region of the osteocalcin promoter. In this report, we show that vitamin D receptors derived from nuclear extracts of mammalian cells bind directly to this cis-acting element in vitro and do so in a manner requiring hormone. Vitamin D receptors derived from reticulocyte lysate translations in vitro or from extracts of a Saccharomyces cerevisiae strain that expresses the recombinant protein also bind the osteocalcin responsive element, but only when nuclear extracts of mammalian cells are provided. The vitamin-D-receptor-DNA-binding accessory factor is isolated by salt extraction, labile to temperature, and sensitive to tryptic digestion. These studies suggest that the high-affinity interaction of the vitamin D receptor with the osteocalcin vitamin D response element in vitro requires both 1,25-dihydroxyvitamin D3 and an accessory protein derived from the mammalian cell nucleus.
Liao
J
J
Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030.
Ozono
K
K
Sone
T
T
McDonnell
D P
DP
Pike
J W
JW
D000818:Animals; D001483:Base Sequence; D002117:Calcitriol / Q000494:pharmacology*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D015345:Oligonucleotide Probes; D015675:Osteocalcin / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid; D015675:Osteocalcin; D004247:DNA; D002117:Calcitriol