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FMIP controls the adipocyte lineage commitment of C2C12 cells by downmodulation of C/EBPalpha

Oncogene volume 26, pages 1020–1027 (2007)Cite this article

Abstract

Fms interacting protein (FMIP) is a substrate for Fms tyrosine kinase, and a nuclear/cytoplasm shuttling protein with a leucine zipper. As the phosphorylation of FMIP is observed in insulin-stimulated preadipocytes, we examined the role of FMIP in adipocyte differentiation, using the mesenchymal multipotent stem cells, C2C12 cells, that can differentiate into adipocytes, muscle cells and osteoblasts. Ectopic expression of FMIP in C2C12 impairs the adipocyte differentiation induced by treatment with insulin, dexamethasone and 3-isobutyl-1-methylxanthine. These cells exhibit muscle phenotype with multinuclear morphology. Furthermore, knockdown of endogenous FMIP expression by small interfering RNA improves adipocytic lineage commitment of C2C12 cells, while impairing muscle differentiation. Upon stimulation with insulin, CCAAT/enhancer binding protein (C/EBP)beta, but not C/EBPalpha, is upregulated in cells expressing ectopic FMIP, whereas in FMIP knockdown cells, C/EBPalpha is constitutively expressed. Ectopic expression of C/EBPalpha counteracts the effects of FMIP, whereas C/EBPalpha knockdown partially mimics the effects of FMIP in this system. Northern blot analysis and reverse transcriptase–polymerase chain reaction study reveal that ectopic FMIP-expressing cells do not contain the polyadenylated C/EBPalpha mRNA, but contain the C/EBPalpha pre-mRNA, suggesting that FMIP plays a role in RNA processing and/or export. Indeed, a member of the THO complex that plays a role in mRNA export, THOC1, is co-precipitated with FMIP. The data we have acquired on FMIP suggest that it is a target for tyrosine kinase receptors that potentiate mRNA export.

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References

  • Cao Z, Umek RM, McKnight SL . (1991). Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev 5: 1538–1552.

    Article  CAS  Google Scholar 

  • Flodby P, Barlow C, Kylefjord H, Ahrlund-Richter L, Xanthopoulos KG . (1996). Increased hepatic cell proliferation and lung abnormalities in mice deficient in CCAAT/enhancer binding protein alpha. J Biol Chem 271: 24753–24760.

    Article  CAS  Google Scholar 

  • Friedman AD . (2002). Transcriptional regulation of granulocyte and monocyte development. Oncogene 21: 3377–3390.

    Article  CAS  Google Scholar 

  • Freytag SO, Paielli DL, Gilbert JD . (1994). Ectopic expression of the CCAAT/enhancer-binding protein alpha promotes the adipogenic program in a variety of mouse fibroblastic cells. Genes Dev 8: 1654–1663.

    Article  CAS  Google Scholar 

  • Fux C, Mitta B, Kramer BP, Fussenegger M . (2004). Dual-regulated expression of C/EBP-alpha and BMP-2 enables differential differentiation of C2C12 cells into adipocytes and osteoblasts. Nucleic Acids Res 32: e1.

    Article  Google Scholar 

  • Gridley S, Lane WS, Garner CW, Lienhard GV . (2005). Novel insulin-elicited phosphoproteins in adipocytes. Cell Signaling 17: 59–66.

    Article  CAS  Google Scholar 

  • Helftenbein G, Krusekopf K, Just U, Cross M, Ostertag W, Niemann H et al. (1996). Transcriptional regulation of the c-fms proto-oncogene mediated by granulocyte/macrophage colony-stimulating factor (GM-CSF) in murine cell lines. Oncogene 12: 931–935.

    CAS  PubMed  Google Scholar 

  • Hu E, Tonzoz 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  CAS  Google Scholar 

  • Johnson PF . (2005). Molecular stop signs: regulation of cell-cycle arrest by C/EBP transcription factors. J Cell Sci 118: 2545–2555.

    Article  CAS  Google Scholar 

  • Jones LC, Lin ML, Chen SS, Klug U, Hofmann WK, Lee S et al. (2002). Expression of C/EBPbeta from the C/ebpalpha gene locus is sufficient for normal hematopoiesis in vivo. Blood 99: 2032–2036.

    Article  CAS  Google Scholar 

  • Kane S, Sano H, Liu SC, Asara JM, Lane WS, Garner CC et al. (2002). A method to identify serine kinase substrates. Akt phosphorylates a novel adipocyte protein with a Rab GTPase-activating protein (GAP) domain. J Biol Chem 277: 22115–22118.

    Article  CAS  Google Scholar 

  • Koch A, Mancini A, El Bounkari O, Tamura T . (2005). The SH2-domian-containing inositol 5-phosphatase (SHIP)-2 binds to c-Met directly via tyrosine residue 1356 and involves hepatocyte growth factor (HGF)-induced lamellipodium formation, cell scattering and cell spreading. Oncogene 24: 3436–3447.

    Article  CAS  Google Scholar 

  • Lane MD, Tang QQ, Jiang MS . (1999). Role of the CCAAT enhancer binding proteins (C/EBPs) in adipocyte differentiation. BBRC 266: 677–683.

    CAS  PubMed  Google Scholar 

  • Lin FT, Lane MD . (1994). CCAAT/enhancer binding protein alpha is sufficient to initiate the 3T3-L1 adipocyte differentiation program. Proc Natl Acad Sci USA 91: 8757–8761.

    Article  CAS  Google Scholar 

  • Mancini A, Koch A, Whetton AD, Tamura T . (2004). The M-CSF receptor substrate and interacting protein FMIP is governed in its subcellular localization by protein kinase C-mediated phosphorylation, and thereby potentiates M-CSF-mediated differentiation. Oncogene 23: 6581–6589.

    Article  CAS  Google Scholar 

  • Mason DY, Cordell JL, Pulford KAF . (1983) In: Bullock GR, Petrusz P (eds). Techniques in Immunocytochemistry. Academic Press: London, pp 175–216.

    Google Scholar 

  • Masuda S, Das R, Cheng H, Hurt E, Dorman N, Reed R . (2005). Recruitment of the human TREX complex to mRNA during splicing. Gene Dev 19: 512–1517.

    Article  Google Scholar 

  • Mori T, Sakaue H, Iguchi H, Gomi H, Okada Y, Takashima Y et al. (2005). Role of Kruppel-like factor 15 (KLF15) in transcriptional regulation of adipogenesis. J Biol Chem 280: 12867–12875.

    Article  CAS  Google Scholar 

  • Otto TC, Lane MD . (2005). Adipose development: from stem cell to adipocyte. Crit Rev Biochem Mol Biol 40: 229–242.

    Article  CAS  Google Scholar 

  • Park BH, Qiang L, Farmer SR . (2004). Phosphorylation of C/EBPbeta at a consensus extracellular signal-regulated kinase/glycogen synthase kinase 3 site is required for the induction of adiponectin gene expression during the differentiation of mouse fibroblasts into adipocytes. Mol Cell Biol 24: 8671–8680.

    Article  CAS  Google Scholar 

  • Popernack PM, Truong LT, Kamphuis M, Henderson AJ . (2001). Ectopic expression of CCAAT/enhancer binding protein beta (C/EBPbeta) in long-term bone marrow cultures induces granulopoiesis and alters stromal cell function. J Hematother Stem Cell Res 10: 631–642.

    Article  CAS  Google Scholar 

  • Rehwinkel J, Herold A, Gari K, Köcher T, Rode M, Ciccarelli FL et al. (2004). Genome-wide analysis of mRNAs regulated by the THO complex in Drosophila melanogaster. Nat Struct Mol Biol 11: 558–566.

    Article  CAS  Google Scholar 

  • Scherr M, Battmer K, Blomer U, Schiedlmeier B, Ganser A, Grez M et al. (2002). Lentiviral gene transfer into peripheral blood-derived CD34+ NOD/SCID-repopulating cells. Blood 99: 709–712.

    Article  CAS  Google Scholar 

  • Scherr M, Battmer K, Ganser A, Eder M . (2003). Modulation of gene expression by lentiviral-mediated delivery of small interfering RNA. Cell Cycle 2: 251–257.

    Article  CAS  Google Scholar 

  • Shi XM, Shi W, Li Q, Song B, Wan M, Bai S et al. (2003). A glucocorticoid-induced leucine-zipper protein. GILZ, inhibits adipogenesis of mesenchymal cells. EMBO Rep 4: 374–380.

    Article  CAS  Google Scholar 

  • Tamura T, Mancini A, Joos H, Koch A, Hakim C, Dumanski J et al. (1999). FMIP, a novel Fms-interacting protein, affects granulocyte/macrophage differentiation. Oncogene 18: 6488–6495.

    Article  CAS  Google Scholar 

  • Tang QQ, Otto TC, Lane MD . (2003). CCAAT/enhancer-binding protein beta is required for mitotic clonal expansion during adipogenesis. Proc Natl Acad Sci USA 100: 850–855.

    Article  CAS  Google Scholar 

  • Tenen DG, Hromas R, Licht JD, Zhang DE . (1997). Transcription factors, normal myeloid development, and leukemia. Blood 90: 489–519.

    CAS  PubMed  Google Scholar 

  • Wang ND, Finegold MJ, Bradley A, Ou CN, Abdelsayed SV, Wilde MD et al. (1995). Impaired energy homeostasis in C/EBP alpha knockout mice. Science 269: 1108–1112.

    Article  CAS  Google Scholar 

  • Wang QF, Friedman AD . (2002). CCAAT/enhancer-binding proteins are required for granulopoiesis independent of their induction of the granulocyte colony-stimulating factor receptor. Blood 99: 2776–2785.

    Article  CAS  Google Scholar 

  • Zhang DE, Zhang P, Wang ND, Hetherington CJ, Darlington GJ, Tenen DG . (1997). Absence of granulocyte colony-stimulating factor signaling and neutrophil development in CCAAT enhancer binding protein alpha-deficient mice. Proc Natl Acad Sci USA 94: 569–574.

    Article  CAS  Google Scholar 

  • Zhang JW, Tang QQ, Vinson C, Lane MD . (2004). Dominant-negative C/EBP disrupts mitotic clonal expansion and differentiation of 3T3-L1 preadipocytes. Proc Natl Acad Sci USA 101: 43–47.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Regina Wilms, Sabine Klebba-Faerber and Iris Dallmann for technical assistance, Renate Scheibe for providing C2C12 cells and Bruce Boschek for critically reading the manuscript. The research was supported by Sonderforschungsbereich566 (B2) and the Deutsche Forschungsgemeinschaft (Ta-111/8/-4). AHB, KP, ADW and LL are supported by the Leukaemia Research Fund and ADW by BBSRC. This work is a part of PhD thesis of A-FN.

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Authors and Affiliations

  1. Institut fuer Biochemie, Medizinische Hochschule Hannover, Hannover, Germany

    A Mancini, O El Bounkari, A-F Norrenbrock & T Tamura

  2. Haematologie und Onkologie, Medizinische Hochschule Hannover, Hannover, Germany

    M Scherr, D Schaefer & M Eder

  3. Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Headington, Oxford, UK

    A H Banham, K Pulford & L Lyne

  4. Cancer Studies Division, Faculty of Medical and Human Sciences, University of Manchester, Christie Hospital, Manchester, UK

    A D Whetton

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  1. A Mancini
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  2. O El Bounkari
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  4. M Scherr
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  10. A D Whetton
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  11. T Tamura
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Correspondence to T Tamura.

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Mancini, A., El Bounkari, O., Norrenbrock, AF. et al. FMIP controls the adipocyte lineage commitment of C2C12 cells by downmodulation of C/EBPalpha. Oncogene 26, 1020–1027 (2007). https://doi.org/10.1038/sj.onc.1209853

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