C/EBPß Reprograms White 3T3-L1 Preadipocytes to a Brown Adipocyte Pattern of Gene Expression

Georgios Karamanlidis, Angeliki Karamitri, Kevin Docherty, David G. Hazlerigg, Michael A. Lomax

Research output: Contribution to journalArticle

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Abstract

cAMP-dependent protein kinase induction of PPAR (gamma) coactivator-1alpha (PGC-1alpha) and uncoupling protein 1 (UCP1) expression is an essential step in the commitment of preadipocytes to the brown adipose tissue (BAT) lineage. We studied the molecular mechanisms responsible for differential expression of PGC-1alpha in HIB1B (BAT) and 3T3-L1 white adipose tissue (WAT) precursor cell lines. In HIB1B cells PGC-1alpha and UCP1 expression is cAMP-inducible, but in 3T3-L1 cells, expression is reduced and is cAMP-insensitive. A proximal 264-bp PGC-1alpha reporter construct was cAMP-inducible only in HIB1B cells and was suppressed by site-directed mutagenesis of the proximal cAMP response element (CRE). In electrophoretic mobility shift assays, the transcription factors CREB and C/EBPß, but not C/EBPa and C/EBPd, bound to the CRE on the PGC-1alpha promoter region in HIB1B and 3T3-L1 cells. Chromatin immunoprecipitation studies demonstrated that C/EBPß and CREB bound to the CRE region in HIB1B and 3T3-L1 cell lysates. C/EBPß expression was induced by cAMP only in HIB1B cells, and overexpression of C/EBPß rescued cAMP-inducible PGC-1alpha and UCP1 expression in 3T3-L1 cells. These data demonstrate that differentiation of preadipocytes toward the BAT rather than the WAT phenotype is controlled in part by the action of C/EBPß on the CRE in PGC-1alpha proximal promoter.

Original languageEnglish
Pages (from-to)24660-24669
Number of pages10
JournalThe Journal of Biological Chemistry
Volume282
Issue number34
DOIs
Publication statusPublished - 21 Jun 2007

Keywords

  • element-binding protein
  • amp response element
  • adipose-tissue
  • transcription factors
  • mice lacking
  • ppar-gamma
  • differentiation
  • coactivator
  • adipogenesis
  • activation

Cite this

Karamanlidis, G., Karamitri, A., Docherty, K., Hazlerigg, D. G., & Lomax, M. A. (2007). C/EBPß Reprograms White 3T3-L1 Preadipocytes to a Brown Adipocyte Pattern of Gene Expression. The Journal of Biological Chemistry, 282(34), 24660-24669. https://doi.org/10.1074/jbc.M703101200

C/EBPß Reprograms White 3T3-L1 Preadipocytes to a Brown Adipocyte Pattern of Gene Expression. / Karamanlidis, Georgios; Karamitri, Angeliki; Docherty, Kevin; Hazlerigg, David G.; Lomax, Michael A.

In: The Journal of Biological Chemistry, Vol. 282, No. 34, 21.06.2007, p. 24660-24669.

Research output: Contribution to journalArticle

Karamanlidis, G, Karamitri, A, Docherty, K, Hazlerigg, DG & Lomax, MA 2007, 'C/EBPß Reprograms White 3T3-L1 Preadipocytes to a Brown Adipocyte Pattern of Gene Expression', The Journal of Biological Chemistry, vol. 282, no. 34, pp. 24660-24669. https://doi.org/10.1074/jbc.M703101200
Karamanlidis, Georgios ; Karamitri, Angeliki ; Docherty, Kevin ; Hazlerigg, David G. ; Lomax, Michael A. / C/EBPß Reprograms White 3T3-L1 Preadipocytes to a Brown Adipocyte Pattern of Gene Expression. In: The Journal of Biological Chemistry. 2007 ; Vol. 282, No. 34. pp. 24660-24669.
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abstract = "cAMP-dependent protein kinase induction of PPAR (gamma) coactivator-1alpha (PGC-1alpha) and uncoupling protein 1 (UCP1) expression is an essential step in the commitment of preadipocytes to the brown adipose tissue (BAT) lineage. We studied the molecular mechanisms responsible for differential expression of PGC-1alpha in HIB1B (BAT) and 3T3-L1 white adipose tissue (WAT) precursor cell lines. In HIB1B cells PGC-1alpha and UCP1 expression is cAMP-inducible, but in 3T3-L1 cells, expression is reduced and is cAMP-insensitive. A proximal 264-bp PGC-1alpha reporter construct was cAMP-inducible only in HIB1B cells and was suppressed by site-directed mutagenesis of the proximal cAMP response element (CRE). In electrophoretic mobility shift assays, the transcription factors CREB and C/EBP{\ss}, but not C/EBPa and C/EBPd, bound to the CRE on the PGC-1alpha promoter region in HIB1B and 3T3-L1 cells. Chromatin immunoprecipitation studies demonstrated that C/EBP{\ss} and CREB bound to the CRE region in HIB1B and 3T3-L1 cell lysates. C/EBP{\ss} expression was induced by cAMP only in HIB1B cells, and overexpression of C/EBP{\ss} rescued cAMP-inducible PGC-1alpha and UCP1 expression in 3T3-L1 cells. These data demonstrate that differentiation of preadipocytes toward the BAT rather than the WAT phenotype is controlled in part by the action of C/EBP{\ss} on the CRE in PGC-1alpha proximal promoter.",
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T1 - C/EBPß Reprograms White 3T3-L1 Preadipocytes to a Brown Adipocyte Pattern of Gene Expression

AU - Karamanlidis, Georgios

AU - Karamitri, Angeliki

AU - Docherty, Kevin

AU - Hazlerigg, David G.

AU - Lomax, Michael A.

PY - 2007/6/21

Y1 - 2007/6/21

N2 - cAMP-dependent protein kinase induction of PPAR (gamma) coactivator-1alpha (PGC-1alpha) and uncoupling protein 1 (UCP1) expression is an essential step in the commitment of preadipocytes to the brown adipose tissue (BAT) lineage. We studied the molecular mechanisms responsible for differential expression of PGC-1alpha in HIB1B (BAT) and 3T3-L1 white adipose tissue (WAT) precursor cell lines. In HIB1B cells PGC-1alpha and UCP1 expression is cAMP-inducible, but in 3T3-L1 cells, expression is reduced and is cAMP-insensitive. A proximal 264-bp PGC-1alpha reporter construct was cAMP-inducible only in HIB1B cells and was suppressed by site-directed mutagenesis of the proximal cAMP response element (CRE). In electrophoretic mobility shift assays, the transcription factors CREB and C/EBPß, but not C/EBPa and C/EBPd, bound to the CRE on the PGC-1alpha promoter region in HIB1B and 3T3-L1 cells. Chromatin immunoprecipitation studies demonstrated that C/EBPß and CREB bound to the CRE region in HIB1B and 3T3-L1 cell lysates. C/EBPß expression was induced by cAMP only in HIB1B cells, and overexpression of C/EBPß rescued cAMP-inducible PGC-1alpha and UCP1 expression in 3T3-L1 cells. These data demonstrate that differentiation of preadipocytes toward the BAT rather than the WAT phenotype is controlled in part by the action of C/EBPß on the CRE in PGC-1alpha proximal promoter.

AB - cAMP-dependent protein kinase induction of PPAR (gamma) coactivator-1alpha (PGC-1alpha) and uncoupling protein 1 (UCP1) expression is an essential step in the commitment of preadipocytes to the brown adipose tissue (BAT) lineage. We studied the molecular mechanisms responsible for differential expression of PGC-1alpha in HIB1B (BAT) and 3T3-L1 white adipose tissue (WAT) precursor cell lines. In HIB1B cells PGC-1alpha and UCP1 expression is cAMP-inducible, but in 3T3-L1 cells, expression is reduced and is cAMP-insensitive. A proximal 264-bp PGC-1alpha reporter construct was cAMP-inducible only in HIB1B cells and was suppressed by site-directed mutagenesis of the proximal cAMP response element (CRE). In electrophoretic mobility shift assays, the transcription factors CREB and C/EBPß, but not C/EBPa and C/EBPd, bound to the CRE on the PGC-1alpha promoter region in HIB1B and 3T3-L1 cells. Chromatin immunoprecipitation studies demonstrated that C/EBPß and CREB bound to the CRE region in HIB1B and 3T3-L1 cell lysates. C/EBPß expression was induced by cAMP only in HIB1B cells, and overexpression of C/EBPß rescued cAMP-inducible PGC-1alpha and UCP1 expression in 3T3-L1 cells. These data demonstrate that differentiation of preadipocytes toward the BAT rather than the WAT phenotype is controlled in part by the action of C/EBPß on the CRE in PGC-1alpha proximal promoter.

KW - element-binding protein

KW - amp response element

KW - adipose-tissue

KW - transcription factors

KW - mice lacking

KW - ppar-gamma

KW - differentiation

KW - coactivator

KW - adipogenesis

KW - activation

U2 - 10.1074/jbc.M703101200

DO - 10.1074/jbc.M703101200

M3 - Article

VL - 282

SP - 24660

EP - 24669

JO - The Journal of Biological Chemistry

JF - The Journal of Biological Chemistry

SN - 0021-9258

IS - 34

ER -