Transcriptional Mechanisms of Bone Morphogenetic Protein-induced Osteoprotegrin Gene Expression

Mei Wan, Xing Ming Shi, Xu Feng, Xu Cao

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Osteoprotegerin (OPG), an osteoblast-secreted decoy receptor, specifically binds to osteoclast differentiation factor and inhibits osteoclast maturation. Members of the transforming growth factor-β superfamily including bone morphogenetic proteins (BMPs) stimulate OPG mRNA expression. In this study, we have characterized the transcription mechanism of BMP-induced OPG gene expression. Transfection of Smad1 and a constitutively active BMP type IA receptor ALK3 (Q233) stimulated the OPG promoter. Deletion analysis of the OPG promoter identified two Hoxc-8 binding sites that respond to BMP stimulation. Glutathione S-transferase-Hoxc-8 protein binds to these two Hox sites specifically. Consistent with the transfection results of the native promoter, ALK3 or Smad1 linker region, which interacts with Hoxc-8, stimulated the activation of the reporter construct with the two Hox sites. Overexpression of Hoxc-8 inhibited the induced promoter activity. When the two Hox binding sites were mutated, ALK3 or Smad1 linker region no longer activated the transcription. Importantly, Smad1 linker region induced both OPG promoter activity and endogenous OPG protein expression in 2T3 osteoblastic cells. The medium from cells transfected with Smad1 linker region expression plasmid effectively inhibited osteoclastogenesis. Collectively, our data indicate that Hox sites mediate both OPG promoter construct activity and endogenous OPG gene expression in response to BMP stimulation.

Original languageEnglish (US)
Pages (from-to)10119-10125
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number13
DOIs
StatePublished - Mar 30 2001
Externally publishedYes

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Osteoprotegerin
Bone Morphogenetic Proteins
Gene expression
Gene Expression
Transcription
Transfection
Type I Bone Morphogenetic Protein Receptors
Binding Sites
RANK Ligand
Osteoblasts
Transforming Growth Factors
Osteoclasts
Glutathione Transferase
Osteogenesis
Proteins
Plasmids
Chemical activation
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Transcriptional Mechanisms of Bone Morphogenetic Protein-induced Osteoprotegrin Gene Expression. / Wan, Mei; Shi, Xing Ming; Feng, Xu; Cao, Xu.

In: Journal of Biological Chemistry, Vol. 276, No. 13, 30.03.2001, p. 10119-10125.

Research output: Contribution to journalArticle

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abstract = "Osteoprotegerin (OPG), an osteoblast-secreted decoy receptor, specifically binds to osteoclast differentiation factor and inhibits osteoclast maturation. Members of the transforming growth factor-β superfamily including bone morphogenetic proteins (BMPs) stimulate OPG mRNA expression. In this study, we have characterized the transcription mechanism of BMP-induced OPG gene expression. Transfection of Smad1 and a constitutively active BMP type IA receptor ALK3 (Q233) stimulated the OPG promoter. Deletion analysis of the OPG promoter identified two Hoxc-8 binding sites that respond to BMP stimulation. Glutathione S-transferase-Hoxc-8 protein binds to these two Hox sites specifically. Consistent with the transfection results of the native promoter, ALK3 or Smad1 linker region, which interacts with Hoxc-8, stimulated the activation of the reporter construct with the two Hox sites. Overexpression of Hoxc-8 inhibited the induced promoter activity. When the two Hox binding sites were mutated, ALK3 or Smad1 linker region no longer activated the transcription. Importantly, Smad1 linker region induced both OPG promoter activity and endogenous OPG protein expression in 2T3 osteoblastic cells. The medium from cells transfected with Smad1 linker region expression plasmid effectively inhibited osteoclastogenesis. Collectively, our data indicate that Hox sites mediate both OPG promoter construct activity and endogenous OPG gene expression in response to BMP stimulation.",
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