The steroid hormone vitamin D is a principal mediator of skeletal homeostasis. 1,25-Dihydroxyvitamin D3 treatment of ROS 17/2.8 osteoblast-like cells results in a ligand-dependent increase in transcription of the bone-specific osteocalcin gene. This transcriptional upregulation requires the positive cis-acting vitamin D responsive element (VDRE). We have used the ligation-mediated polymerase chain reaction to demonstrate that protein occupancy of the VDRE within the intact cell correlates with increased synthesis of osteocalcin transcripts. These protein-DNA contacts were not present in the absence of vitamin D or in osteosarcoma cells (ROS 24.1) lacking the vitamin D receptor. Our results establish in intact cells the requirement for both ligand- and receptor-dependent occupancy of the VDRE for vitamin D responsive enhancement of osteocalcin gene transcription.
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Animals
Base Sequence
Binding Sites
Calcitriol
DNA-Binding Proteins
Gene Expression Regulation
In Vitro Techniques
Molecular Sequence Data
Oligodeoxyribonucleotides
Osteocalcin
Osteosarcoma
Promoter Regions, Genetic
RNA, Messenger
Rats
Receptors, Calcitriol
Transcription, Genetic
Tumor Cells, Cultured
