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Journal of Dental Research
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Biological

Phosphate Regulates Osteopontin Gene Transcription

S. Fatherazi1, D. Matsa-Dunn2, B.L. Foster1,2, R.B. Rutherford1, M.J. Somerman1,2 and R.B. Presland1,3,*

1 Departments of Oral Biology and
2 Periodontics, School of Dentistry, Box 357132, and
3 Department of Medicine (Dermatology), University of Washington, Seattle, WA 98195-7132, USA

Correspondence: * corresponding author, rp{at}u.washington.edu

Extracellular inorganic phosphate (ePi) is a key regulator of cementoblast behavior, both in vivo and in vitro, and results in a marked increase in osteopontin expression in vitro. To examine the molecular mechanisms involved in ePi induction of osteopontin gene expression, we transfected a series of osteopontin promoter-luciferase constructs into OCCM-30 cementoblasts. Our results demonstrate that ePi can directly induce osteopontin gene transcription. The region responsive to ePi signaling was localized to a 53-bp region of the promoter between –1454 and –1401 that contains a glucocorticoid response element (GRE). Mutation of the GRE abolished the ePi response, suggesting that glucocorticoid receptor (GR) signaling is required for ePi-mediated transcription. In addition, treatment of cells with the GR antagonist RU-486 (Mifepristone) prevented promoter activation by ePi. The results presented support a model demonstrating that inorganic phosphate regulates OPN gene transcription in cementoblasts through a pathway that requires a functional GR.

Key Words: phosphate • osteopontin • gene transcription • BSP, bone sialoprotein • DMP1, dentin matrix protein 1 • EMSA, Electrophoretic mobility shift assay • ePi, extracellular inorganic phosphate • GRE, glucocorticoid response element • GR, glucocorticoid receptor • OPN, osteopontin • Pi, inorganic phosphate • Q-PCR, quantitative PCR.

Journal of Dental Research, Vol. 88, No. 1, 39-44 (2009)
DOI: 10.1177/0022034508328072
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