Vitamin D-mediated modifications in protein-DNA interactions at two promoter elements of the osterocalcin gene

Journal Article · Thu Mar 01 04:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.5.1701· OSTI ID:6153022

Markose, E R; Stein, J L; Stein, G S; Lian, J B ^[1]

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1. Univ. of Massachusetts Medical Center, Worcester (USA)

By the combined use of DNase I footprinting, electrophoretic mobility-shift assay, and methylation interference analysis, the authors have identified a series of sequence-specific protein-DNA interactions in the 5{prime} flanking region of the rat osteocalcin gene. Stimulation of osteocalcin gene expression by 1,25-dihydroxyvitamin D{sub 3}, a physiologic mediator of this bone-specific gene in vitro and in vivo, is associated with modifications in the binding of ROS 17/2.8 cell nuclear factors to two promoter segments that up-regulate transcription. One segment located between -462 and -437 exhibits a vitamin D-dependent increase in sequence-specific binding of nuclear factors. This element (CTGGGTGAATGAGGACATTACT-GACC), identified at single nucleotide resolution, contains a region of hyphenated dyad symmetry and shares sequence homology with consensus steroid-responsive elements and with the sequence that has been identified as the vitamin D receptor binding site in the human osteocalcin gene. They have also observed that vitamin D stimulation of osteocalcin gene expression results in a 5-fold increase in protein binding to the region of the osteocalcin box, a 24-nucleotide segment in the proximal promoter with a CCAAT motif as the central core. The results demonstrate protein-DNA interactions in a vitamin D-responsive element and in a second sequence, the osteocalcin box, both of which are involved in the physiologic regulation of the osteocalcin gene in response to 1,25-dihydroxyvitamin D{sub 3}.

• OSTI ID: 6153022
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 87:5; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
ANIMALS
AUTORADIOGRAPHY
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL FUNCTIONS
BODY
BONE TISSUES
CONFORMATIONAL CHANGES
CONNECTIVE TISSUE
DNA SEQUENCING
DNA-ASE
ENZYMES
ESTERASES
FUNCTIONS
GENE OPERONS
GENE REGULATION
GENE REPRESSORS
HYDROLASES
KINETICS
MAMMALS
MESSENGER-RNA
NUCLEIC ACIDS
NUCLEOPROTEINS
ONTOGENESIS
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PROTEINS
RATS
REACTION KINETICS
RNA
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
TISSUES
TRANSCRIPTION FACTORS
VERTEBRATES
VITAMIN D
VITAMINS