Differential regulation of dentin sialophosphoprotein expression by Runx2 during odontoblast cytodifferentiation.
Dentin sialophosphoprotein (DSPP) consists of dentin sialoprotein (DSP) and dentin phosphoprotein (DPP). The spatial-temporal expression of DSPP is largely restricted during differentiational stages of dental cells. DSPP plays a vital role in tooth development. It is known that an osteoblast-specific transcription factor, Runx2, is essential for osteoblast differentiation. However, effects of Runx2 on DSPP transcription remain unknown. Here, we studied different roles of Runx2 in controlling DSPP expression in mouse preodontoblast (MD10-F2) and odontoblast (MO6-G3) cells. Two Runx2 isoforms were expressed in preodontoblast and odontoblast cells, and in situ hybridization assay showed that DSPP expression increased, whereas Runx2 was down-regulated during odontoblast differentiation and maturation. Three potential Runx2 sites are present in promoters of mouse and rat DSPP genes. Runx2 binds to these sites as demonstrated by electrophoretic mobility shift assay and supershift experiments. Mutations of Runx2 sites in mouse DSPP promoter resulted in a decline of promoter activity in MD10-F2 cells compared with an increase of its activity in MO6-G3 cells. Multiple Runx2 sites were more active than a single site in regulating the DSPP promoter. Furthermore, forced overexpression of Runx2 isoforms induced increases of endogenous DSPP protein levels in MD10-F2 cells but reduced its expression in MO6-G3 cells consistent with the DSPP promoter analysis. Thus, our results suggest that differential positive and negative regulation of DSPP by Runx2 is dependent on use of cytodifferentiation of dental ectomesenchymal-derived cells that may contribute to the spatial-temporal expression of DSPP during tooth development.
Department of Pediatric Dentistry, The University of Texas Health Science Center, San Antonio, 78229-3900, USA. hyuan@emory.edu
Shuo
S
Chen
Sheela
S
Rani
Yimin
Y
Wu
Aaron
A
Unterbrink
Ting Ting
TT
Gu
Jelica
J
Gluhak-Heinrich
Hui-Hsiu
HH
Chuang
Mary
M
Macdougall
D000818:Animals; D002113:Calcification, Physiologic; D002454:Cell Differentiation; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit; D004268:DNA-Binding Proteins / Q000502:physiology*; D016326:Extracellular Matrix Proteins; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D051379:Mice; D009804:Odontoblasts / Q000166:cytology*; D010750:Phosphoproteins; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics*; D012795:Sialoglycoproteins; D014070:Tooth / Q000254:growth & development; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000502:physiology*; D000099311:Dentin Sialophosphoprotein
D050659:Core Binding Factor Alpha 1 Subunit; D004268:DNA-Binding Proteins; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D011498:Protein Precursors; D012795:Sialoglycoproteins; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D000099311:Dentin Sialophosphoprotein; C493716:Runx2 protein, mouse