Abstract
The zinc finger-containing transcription factors Egr1 (Krox24) and Egr2 (Krox20) have been implicated in the proliferation and differentiation of many cell types. Egr2 has earlier been shown to play a positive role in adipocyte differentiation, but the function of Egr1 in this context is unknown. We compared the roles of Egr1 and Egr2 in the differentiation of murine 3T3-L1 adipocytes. Egr1 protein was rapidly induced after addition of differentiation cocktail, whereas Egr2 protein initially remained low before increasing on days 1 and 2, concomitant with the disappearance of Egr1. In marked contrast to the effects of Egr2, differentiation was inhibited by ectopic expression of Egr1 and potentiated by knockdown of Egr1. The pro-adipogenic effects of Egr1 knockdown were particularly notable when isobutylmethylxanthine (IBMX) was omitted from the differentiation medium. However, knockdown of Egr1 did not affect CCAAT/enhancer binding protein (C/EBP)β protein expression or phosphorylation of CREB Ser133. Further, Egr1 did not directly affect the activity of promoters for the master adipogenic transcription factors, C/EBPα or peroxisome proliferator-activated receptor-γ2, as assessed in luciferase reporter assays. These data indicate that Egr1 and Egr2 exert opposing influences on adipocyte differentiation and that the careful regulation of both is required for maintaining appropriate levels of adipogenesis. Further, the pro-differentiation effects of IBMX involve suppression of the inhibitory influence of Egr1.
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Abbreviations
- aP2:
-
fatty-acid binding protein 2
- C/EBP:
-
CCAAT/enhancer binding protein
- DMEM:
-
Dulbecco's modified Eagle's medium
- GLUT4:
-
glucose transporter 4
- HTNC:
-
His-TAT-NLS-Cre recombinase fusion protein
- IBMX:
-
isobutylmethylxanthine
- IPTG:
-
isopropyl β-D-thiogalactoside
- KLF:
-
Kruppel-like factor
- MDI:
-
IBMX-dexamethasone and insulin
- PBS:
-
phosphate buffered saline
- PTB:
-
phosphate Tris buffer
- PPAR:
-
peroxisome proliferator-activated receptor
- SDS-PAGE:
-
sodium dodecyl sulfate polyacrylamide gel electrophoresis
- RT-PCR:
-
reverse transcriptase polymerase chain reaction
- shRNA:
-
short hairpin ribonucleic acid
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Acknowledgements
We are very grateful to the Wellcome Trust for financial support for the Biological Atlas of Insulin Resistance (BAIR) program (GR066786), and to all members of the BAIR consortium for helpful discussion. We also acknowledge support from the BBSRC (project grant C20109 for SHR), MRC (studentships for SV and WPC) and European Union COST Action BM0602. We thank Dr. F Edenhofer (Institute of Reconstructive Neurobiology, University of Bonn, Germany) for the pTriEx1.1 plasmid and advice on the production of HTNC protein, Dr. MJ Birnbaum (Howard Hughes Medical Institute, PA, USA) for pLPNPX1 plasmid and Dr. Justin Rochford (University of Cambridge) for helpful discussion.
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Boyle, K., Hadaschik, D., Virtue, S. et al. The transcription factors Egr1 and Egr2 have opposing influences on adipocyte differentiation. Cell Death Differ 16, 782–789 (2009). https://doi.org/10.1038/cdd.2009.11
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DOI: https://doi.org/10.1038/cdd.2009.11
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