Tandem repeat of C/EBP binding sites mediates PPARγ2 gene transcription in glucocorticoid-induced adipocyte differentiation

Shi Xing Ming, Harry C. Blair, Xiangli Yang, Jay M. McDonald, Xu Cao

Research output: Contribution to journalArticle

Abstract

Bone marrow stromal stem cells differentiate into many different types of cells including osteoblasts and adipocytes. Long-term glucocorticoid treatment decreases osteoblastic activity but increases adipocytes. We investigated the mechanism of glucocorticoid-induced PPARγ2 transcription. Treatment of human bone marrow stromal cells with dexamethasone induced the differentiation of these cells into adipocytes as measured by oil-red O staining, and Northern blot analysis showed that dexamethasone strongly induced PPARγ2 mRNA expression in cells cultured in adipocyte induction medium. Moreover, the mRNA of C/EBPδ, an adipocyte-promoting transcription factor, was also induced by dexamethasone in the presence of induction medium. Gel mobility shift assays using purified GST-C/EBPδ fusion protein showed that C/EBPδ specifically binds to a 40-base pair DNA element from PPARγ2 promoter, which was found to contain a tandem repeat of C/EBP binding sites. Transfection studies in mouse mesenchymal C3H10T1/2 cells showed that it is the tandem repeat of the C/EBP binding site in PPARγ2 promoter region that regulates dexamethasone-mediated PPARγ2 gene activation. We conclude that glucocorticoid-induced adipogenesis from bone marrow stromal cells is mediated through a reaction cascade in which dexamethasone transcriptionally activates C/EBPδ; C/EBPδ then binds to PPARγ2 promoter and transactivates PPARγ2 gene expression. This activated master regulator, in turn, initiates the adipocyte differentiation.

Original languageEnglish (US)
Pages (from-to)518-527
Number of pages10
JournalJournal of Cellular Biochemistry
Volume76
Issue number3
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Tandem Repeat Sequences
Transcription
Adipocytes
Dexamethasone
Glucocorticoids
Genes
Binding Sites
Mesenchymal Stromal Cells
Bone
CCAAT-Enhancer-Binding Proteins
Messenger RNA
Osteoblasts
Adipogenesis
Stem cells
Genetic Promoter Regions
Gene expression
Electrophoretic Mobility Shift Assay
Protein C
Assays
Base Pairing

Keywords

  • Adipocyte
  • Bone marrow stromal cell
  • Glucocorticoid
  • Osteoblast
  • Osteoporosis

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Tandem repeat of C/EBP binding sites mediates PPARγ2 gene transcription in glucocorticoid-induced adipocyte differentiation. / Xing Ming, Shi; Blair, Harry C.; Yang, Xiangli; McDonald, Jay M.; Cao, Xu.

In: Journal of Cellular Biochemistry, Vol. 76, No. 3, 2000, p. 518-527.

Research output: Contribution to journalArticle

Xing Ming, Shi ; Blair, Harry C. ; Yang, Xiangli ; McDonald, Jay M. ; Cao, Xu. / Tandem repeat of C/EBP binding sites mediates PPARγ2 gene transcription in glucocorticoid-induced adipocyte differentiation. In: Journal of Cellular Biochemistry. 2000 ; Vol. 76, No. 3. pp. 518-527.
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AB - Bone marrow stromal stem cells differentiate into many different types of cells including osteoblasts and adipocytes. Long-term glucocorticoid treatment decreases osteoblastic activity but increases adipocytes. We investigated the mechanism of glucocorticoid-induced PPARγ2 transcription. Treatment of human bone marrow stromal cells with dexamethasone induced the differentiation of these cells into adipocytes as measured by oil-red O staining, and Northern blot analysis showed that dexamethasone strongly induced PPARγ2 mRNA expression in cells cultured in adipocyte induction medium. Moreover, the mRNA of C/EBPδ, an adipocyte-promoting transcription factor, was also induced by dexamethasone in the presence of induction medium. Gel mobility shift assays using purified GST-C/EBPδ fusion protein showed that C/EBPδ specifically binds to a 40-base pair DNA element from PPARγ2 promoter, which was found to contain a tandem repeat of C/EBP binding sites. Transfection studies in mouse mesenchymal C3H10T1/2 cells showed that it is the tandem repeat of the C/EBP binding site in PPARγ2 promoter region that regulates dexamethasone-mediated PPARγ2 gene activation. We conclude that glucocorticoid-induced adipogenesis from bone marrow stromal cells is mediated through a reaction cascade in which dexamethasone transcriptionally activates C/EBPδ; C/EBPδ then binds to PPARγ2 promoter and transactivates PPARγ2 gene expression. This activated master regulator, in turn, initiates the adipocyte differentiation.

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