Spatial expression of Cbfa1/Runx2 isoforms in teeth and characterization of bindings sites in the DSPP gene

Shuo Chen, Ting Ting Gu, Taduru Sreenath, Ashok B. Kulkarni, Gerard Karsenty, Mary MacDougall

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Cbfa1/Runx2 is an essential transcription factor for osteoblast and odontoblast differentiation. Heterogeneous mutations of Cbfa1 gene result in cleidocranial dysplasia, an autosomal dominant syndrome, characterized by abnormal skeletal genesis and dental disorders. Recently three Cbfa1/Runx isoforms (Pebp2αA/type I, til-1/type II, and Osf2/type III) have been identified that differ in their amino-terminal sequences. The precise roles of Cbfa1/Runx2 isoforms in odontoblast development are not known. The purpose of this study was to determine and compare expression patterns of the three Cbfa1/Runx2 isoforms in newborn tooth organs. Toward this aim, we developed three probes: type I and type II, which specifically hybridize with Pebp2αA and til-1, respectively, and type II/III, which hybridizes with osf2 and partially with til-1. In addition, Cbfa1/Runx2 binding sites were identified in the regulatory elements of mouse dentin sialophosphoprotein (mDSPP) gene, which encodes a matrix protein expressed during odontogenesis. In situ hybridization performed with the specific Cbfa1/Runx2 isoform probes demonstrated that all isoforms are expressed in teeth and bone. The type I isoform was expressed at higher levels than isoforms type II and type II/III in developing newborn mouse incisors. Genomic mDSPP clones were isolated and characterized containing ∼2.6 kb of the promoter region. Computer analysis of the promoter segment and intron 1 revealed a number of potential transcriptional factor binding sites including five Cbfa1/Runx2 binding sites, three in the promoter region and two within intron 1. DNA-protein assay and antibody supershift experiments showed that these binding sites interact with nuclear extracts isolated from the mouse odontoblast cell line MO6-G3. Further characterization of the functional role of Cbfa1/Runx2 in the regulation of the mDSPP gene expression is being investigated.

Original languageEnglish (US)
Pages (from-to)338-344
Number of pages7
JournalConnective Tissue Research
Volume43
Issue number2-3
StatePublished - 2002

Fingerprint

Tooth
Protein Isoforms
Genes
Binding Sites
Odontoblasts
Genetic Promoter Regions
Introns
Odontogenesis
Cleidocranial Dysplasia
Osteoblasts
Incisor
Gene expression
In Situ Hybridization
Assays
Bone
Proteins
Transcription Factors
Clone Cells
Cells
Gene Expression

Keywords

  • Cbfa1/Runx2 isoforms
  • Dentin sialophosphoprotein
  • Gene regulation
  • In situ hybridization
  • Odontogenesis

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Nephrology
  • Cell Biology
  • Orthopedics and Sports Medicine

Cite this

Chen, S., Gu, T. T., Sreenath, T., Kulkarni, A. B., Karsenty, G., & MacDougall, M. (2002). Spatial expression of Cbfa1/Runx2 isoforms in teeth and characterization of bindings sites in the DSPP gene. Connective Tissue Research, 43(2-3), 338-344.

Spatial expression of Cbfa1/Runx2 isoforms in teeth and characterization of bindings sites in the DSPP gene. / Chen, Shuo; Gu, Ting Ting; Sreenath, Taduru; Kulkarni, Ashok B.; Karsenty, Gerard; MacDougall, Mary.

In: Connective Tissue Research, Vol. 43, No. 2-3, 2002, p. 338-344.

Research output: Contribution to journalArticle

Chen, S, Gu, TT, Sreenath, T, Kulkarni, AB, Karsenty, G & MacDougall, M 2002, 'Spatial expression of Cbfa1/Runx2 isoforms in teeth and characterization of bindings sites in the DSPP gene', Connective Tissue Research, vol. 43, no. 2-3, pp. 338-344.
Chen, Shuo ; Gu, Ting Ting ; Sreenath, Taduru ; Kulkarni, Ashok B. ; Karsenty, Gerard ; MacDougall, Mary. / Spatial expression of Cbfa1/Runx2 isoforms in teeth and characterization of bindings sites in the DSPP gene. In: Connective Tissue Research. 2002 ; Vol. 43, No. 2-3. pp. 338-344.
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abstract = "Cbfa1/Runx2 is an essential transcription factor for osteoblast and odontoblast differentiation. Heterogeneous mutations of Cbfa1 gene result in cleidocranial dysplasia, an autosomal dominant syndrome, characterized by abnormal skeletal genesis and dental disorders. Recently three Cbfa1/Runx isoforms (Pebp2αA/type I, til-1/type II, and Osf2/type III) have been identified that differ in their amino-terminal sequences. The precise roles of Cbfa1/Runx2 isoforms in odontoblast development are not known. The purpose of this study was to determine and compare expression patterns of the three Cbfa1/Runx2 isoforms in newborn tooth organs. Toward this aim, we developed three probes: type I and type II, which specifically hybridize with Pebp2αA and til-1, respectively, and type II/III, which hybridizes with osf2 and partially with til-1. In addition, Cbfa1/Runx2 binding sites were identified in the regulatory elements of mouse dentin sialophosphoprotein (mDSPP) gene, which encodes a matrix protein expressed during odontogenesis. In situ hybridization performed with the specific Cbfa1/Runx2 isoform probes demonstrated that all isoforms are expressed in teeth and bone. The type I isoform was expressed at higher levels than isoforms type II and type II/III in developing newborn mouse incisors. Genomic mDSPP clones were isolated and characterized containing ∼2.6 kb of the promoter region. Computer analysis of the promoter segment and intron 1 revealed a number of potential transcriptional factor binding sites including five Cbfa1/Runx2 binding sites, three in the promoter region and two within intron 1. DNA-protein assay and antibody supershift experiments showed that these binding sites interact with nuclear extracts isolated from the mouse odontoblast cell line MO6-G3. Further characterization of the functional role of Cbfa1/Runx2 in the regulation of the mDSPP gene expression is being investigated.",
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