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Transcriptional coactivator EDF-1 is required for PPARγ-stimulated adipogenesis

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Abstract

Peroxisome proliferator-activated receptor-γ (PPARγ) is essential for adipogenesis. Since EDF-1 is a cofactor of PPARγ, we investigated the molecular cross-talk between EDF-1 and PPARγ in adipogenesis. While EDF-1 was not modulated during differentiation of 3T3-L1 cells, it co-immunoprecipitated with PPARγ. Silencing EDF-1 by shRNAs inhibited the differentiation in adipocytes of 3T3-L1 cells, as detected by the staining of intracellular triglycerides and the expression of the PPARγ target gene aP2. Accordingly, we found that anti-EDF-1 shRNAs decreased ligand dependent activation of PPARγ in 3T3-L1 transiently transfected with a vector expressing luciferase under the control of a PPARγ responsive consensus. To rule out that this inhibition is due to the concomitant downregulation of PPARγ levels, we overexpressed PPARγ in 3T3-L1 silencing EDF-1 and found a decrease of ligand dependent activation of PPARγ, in spite of the high amounts of PPARγ. These results demonstrate that EDF-1 is required for PPARγ transcriptional activation during 3T3-L1 differentiation.

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References

  1. Rosen ED, Walkey CJ, Puigserver P, Spiegelman BM (2000) Transcriptional regulation of adipogenesis. Genes Dev 14:1293–1307

    PubMed  CAS  Google Scholar 

  2. Gurnell M (2005) Peroxisome proliferator-activated receptor gamma and the regulation of adipocyte function: lessons from human genetic studies. Best Pract Res Clin Endocrinol Metab 19:501–523

    Article  PubMed  CAS  Google Scholar 

  3. Knouff C, Auwerx J (2004) Peroxisome proliferator-activated receptor-gamma calls for activation in moderation: lessons from genetics and pharmacology. Endocr Rev 25:899–918

    Article  PubMed  CAS  Google Scholar 

  4. Evans RM, Barish GD, Wang YX (2004) PPARs and the complex journey to obesity. Nat Med 10:355–361

    Article  PubMed  CAS  Google Scholar 

  5. Rosen ED, Spiegelman BM (2001) PPARgamma: a nuclear regulator of metabolism, differentiation, and cell growth. J Biol Chem 276:37731–37734

    Article  PubMed  CAS  Google Scholar 

  6. Lehrke M, Lazar MA (2005) The many faces of PPARgamma. Cell 123:993–999

    Article  PubMed  CAS  Google Scholar 

  7. Rosenfeld MG, Lunyak VV, Glass CK (2005) Sensors and signals: a coactivator/corepressor/epigenetic code for integrating signal-dependent programs of transcriptional response. Genes Dev 20:1405–1428

    Article  Google Scholar 

  8. Brendel C, Gelman L, Auwerx J (2002) Multiprotein bridging factor-1 (MBF-1) is a cofactor for nuclear receptors that regulate lipid metabolism. Mol Endocrinol 16:1367–1377

    Article  PubMed  CAS  Google Scholar 

  9. Dragoni I, Mariotti M, Consalez G, Soria M, Maier JAM (1998) EDF-1, a novel gene product downregulated in human endothelial cell differentiation. J Biol Chem 273:31119–31124

    Article  PubMed  CAS  Google Scholar 

  10. Mariotti M, Avon E, De Benedictis L, Maier JAM (2000) Interaction between endothelial differentiation-related factor-1 and calmodulin in vitro and in vivo. J Biol Chem 275:24047–24051

    Article  PubMed  CAS  Google Scholar 

  11. Ballabio E, Mariotti M, De Benedictis L, Maier JAM (2004) The dual role of EDF-1 in the cytosol and in the nucleus: modulation by PKA. Mol Cell Life Sci 61:1069–1074

    Article  CAS  Google Scholar 

  12. Bernardini D, Mariotti M, Ballabio E, Maier JAM (2005) Differential expression of EDF-1 and endothelial nitric oxide synthase by proliferating, quiescent and senescent microvascular endothelial cells. Biochim Biophys Acta Mol Cell Res 1745:265–272

    Article  CAS  Google Scholar 

  13. Mishima M, Ozaki J, Ikegami T, Kabe Y, Goto M, Ueda H, Hirose S, Handa H, Shirakawa M (1999) Resonance assignments, secondary structure and 15 N relaxation data of the human transcriptional coactivator hMBF1 (57–148). J Biomol NMR 14:373–376

    Article  PubMed  CAS  Google Scholar 

  14. Miotto B, Struhl K (2006) Differential gene regulation by selective association of transcriptional coactivators and bZIP DNA-binding domains. Mol Cell Biol 26:5969–5982

    Article  PubMed  CAS  Google Scholar 

  15. Stephens JM, Lee J, Pilch PF (1997) Tumor necrosis factor-alpha-induced insulin resistance in 3T3–L1 adipocytes is accompanied by a loss of insulin receptor substrate-1 and GLUT4 expression without a loss of insulin receptor-mediated signal transduction. J Biol Chem 272:971–976

    Article  PubMed  CAS  Google Scholar 

  16. Tzameli I, Fang H, Ollero M, Shi H, Hamm JK, Kievit P, Hollenberg AN, Flier JS (2004) Regulated production of a peroxisome-proliferator activated receptor-γ ligand during an early phase of adipocyte differentiation in 3T3–L1 adipocyte. J Biol Chem 279:1093–1102

    Article  Google Scholar 

  17. Sentinelli F, Filippi E, Cavallo MG, Romeo S, Fanelli M, Baroni MG (2006) The G972R variant of the insulin receptor substrate-1 gene impairs insulin signaling and cell differentiation in 3T3–L1 adipocytes; treatment with a PPARgamma agonist restores normal cell signaling and differentiation. J Endocrinol 188(2):271–285

    Article  PubMed  CAS  Google Scholar 

  18. Metzger D, Imai T, Jiang M, Takukawa R, Desvergne B, Wahli W, Chambon P (2005) Functional role of RXRs and PPAR gamma in mature adipocytes. Prostaglandins Leukot Essent Fatty Acids 73(1):51–58

    Article  PubMed  CAS  Google Scholar 

  19. Takemaru K, Li FQ, Ueda H, Hirose S (1997) Multiprotein bridging factor 1 (MBF1) is an evolutionarily conserved transcriptional coactivator that connects a regulatory factor and TATA element-binding protein. Proc Natl Acad Sci USA 94:7251–7256

    Article  PubMed  CAS  Google Scholar 

  20. Wu Z, Xie Y, Bucher NL, Farmer SR (1995) Conditional ectopic expression of C/EBP beta in NIH-3T3 cells induces PPAR gamma and stimulates adipogenesis. Genes Dev 9:2350–2363

    Article  PubMed  CAS  Google Scholar 

  21. Hu E, Tontonoz P, Spiegelman BM (1995) Transdifferentiation of myoblasts by the adipogenic transcription factors PPAR gamma and C/EBP alpha. Proc Natl Acad Sci USA 92:9856–9860

    Article  PubMed  CAS  Google Scholar 

  22. Albright SR, Tjian R (2000) TAFs revisited: more data reveal new twists and confirm old ideas. Gene 242:1–13

    Article  PubMed  CAS  Google Scholar 

  23. Fox KE, Fankell DM, Erickson PF, Majka SM, Crossno JT Jr, Klemm DJ (2006) Depletion of cAMP-response element-binding protein/ATF1 inhibits adipogenic conversion of 3T3–L1 cells ectopically expressing CCAAT/enhancer-binding protein (C/EBP) alpha, C/EBP beta, or PPAR gamma 2. J Biol Chem 281:40341–40353

    Article  PubMed  CAS  Google Scholar 

  24. Takahashi Y, Inoue J, Kagechika H, Sato R (2009) ApoC-III gene expression is sharply increased during adipogenesis and is augmented by retinoid X receptor (RXR) agonists. FEBS Lett 583:493–497

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Preclinical Sciences, Università di Milano Medical School, Via GB Grassi 74, Milan, Italy

    Marzia Leidi, Massimo Mariotti & Jeanette A. M. Maier

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  1. Marzia Leidi
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  2. Massimo Mariotti
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  3. Jeanette A. M. Maier
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Correspondence to Jeanette A. M. Maier.

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M. Leidi and M. Mariotti have contributed equally to this work.

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Leidi, M., Mariotti, M. & Maier, J.A.M. Transcriptional coactivator EDF-1 is required for PPARγ-stimulated adipogenesis. Cell. Mol. Life Sci. 66, 2733–2742 (2009). https://doi.org/10.1007/s00018-009-0069-4

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  • DOI: https://doi.org/10.1007/s00018-009-0069-4

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