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J. Biol. Chem., Vol. 276, Issue 8, 5459-5466, February 23, 2001

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Identification of a Novel Response Element in the Rat Bone Sialoprotein (BSP) Gene Promoter that Mediates Constitutive and Fibroblast Growth Factor 2-induced Expression of BSP^*

Emi Shimizu-Sasaki, Muneyoshi Yamazaki, Shunsuke Furuyama^§, Hiroshi Sugiya^§, Jaro Sodek^¶, and Yorimasa Ogata^**

From the Departments of  Endodontics, ^§ Physiology, and  Periodontology, Nihon University School of Dentistry at Matsudo, Chiba 271-8587, Japan and the ^¶ Canadian Institutes of Health Research Group in Periodontal Physiology, Faculty of Dentistry and Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, Ontario M5S 3E2, Canada

Bone sialoprotein (BSP) is a sulfated and phosphorylated glycoprotein, found almost exclusively in mineralized connective tissues, that may function in the nucleation of hydroxyapatite crystals. We have found that expression of BSP in osteoblastic ROS 17/2.8 cells is stimulated by fibroblast growth factor 2 (FGF2), a potent mitogen for mesenchymal cells. Stimulation of BSP mRNA with 10 ng/ml FGF2 was first evident at 3 h (~2.6-fold) and reached maximal levels at 6 h (~4-fold). From transient transfection assays, a FGF response element (FRE) was identified (nucleotides 92 to 85, "GGTGAGAA") as a target of transcriptional activation by FGF2. Ligation of two copies of the FRE 5' to an SV40 promoter was sufficient to confer FGF-responsive transcription. A sequence-specific protein-DNA complex, formed with a double-stranded oligonucleotide encompassing the FRE and nuclear extracts from ROS 17/2.8 cells, but not from fibroblasts, was increased following FGF2 stimulation. Several point mutations within the critical FRE sequence abrogated the formation of this complex and suppressed both basal and FGF2-mediated promoter activity. These studies, therefore, have identified a novel FRE in the proximal promoter of the BSP gene that mediates both constitutive and FGF2-induced BSP transcription.

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