Identification of a DNA sequence responsible for binding of the 1,25-dihydroxyvitamin D sub 3 receptor and 1,25-dihydroxyvitamin D sub 3 enhancement of mouse secreted phosphoprotein 1 (Spp-1 or osteopontin) gene expression

Journal Article · Sat Dec 01 00:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.87.24.9995· OSTI ID:5079772

Noda, Masaki; Vogel, R L ^[1]; Craig, A M ^[2]; Prahl, J; DeLuca, H F ^[3]; Denhardt, D T ^[4]

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1. Merck Sharp and Dohme Research Laboratories, West Point, PA (United States)
2. National Inst. of Health, Bethesda, MD (United States)
3. Univ. of Wisconsin, Madison (United States)
4. Rutgers Univ., Piscataway, NJ (United States)

Secreted phosphoprotein 1 (Spp-1; osteopontin) is one of the abundant noncollagenous proteins in bone matrix and is produced by osteoblasts. The authors examined the promoter region of the mouse Spp-1 gene and identified a sequence responsible for 1,25-dihydroxyvitamin D{sub 3} enhancement of the Spp-1 gene expression. This 24-base-pair (bp) sequence (vitamin D response element) is located 761 bp upstream of the transcription start site and consists of two direct repeats of a unique 9-bp motif, AGGTTCACG. The vitamin D response element confers responsiveness of a heterologous promoter to 1,25-dihydroxyvitamin D{sub 3} in a position-and orientation-independent and copy-number-dependent manner. The basal level of expression of the reporter constructs containing this sequence and its response to 1,25-dihydroxyvitamin D{sub 3} were not affected by cotreatment with transforming growth factor {beta} or the tumor promoter phorbol 12-myristate 13-acetate or by cotransfection with a JUN expression vector. The vitamin D response element forms DNA-protein complexes, as indicated by gel-retardation assays. The addition of a monoclonal antibody raised against the vitamin D receptor further retarded the mobility of the DNA-protein complex. Another antibody that recognized the DNA binding region of the vitamin D receptor attenuated its binding to the sequence. These results indicate that this 24-bp sequence containing two 9-bp motifs binds to the vitamin D receptor and mediates the vitamin D{sub 3} enhancement of murine Spp-1 gene expression.

• OSTI ID: 5079772
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 87:24; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
BONE TISSUES
BIOCHEMISTRY
GENE REPRESSORS
DNA SEQUENCING
PHOSPHOPROTEINS
SECRETION
MICE
MONOCLONAL ANTIBODIES
RECEPTORS
VITAMIN D
ANIMAL TISSUES
ANIMALS
ANTIBODIES
BODY
CHEMISTRY
CONNECTIVE TISSUE
MAMMALS
MEMBRANE PROTEINS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PROTEINS
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
TISSUES
VERTEBRATES
VITAMINS
550201* - Biochemistry- Tracer Techniques