Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Analysis of cell cycle arrest in adipocyte differentiation

Oncogene volume 18, pages 459–466 (1999)Cite this article

Abstract

Confluent 3T3-L1 preadipocytes differentiate to adipocytes in the presence of insulin, dexamethasone, and isobutylmethylxanthine (IDI). A transient increase of DNA synthesis is induced in 3T3-L1 cells 18 h after addition of IDI, followed by an arrest in the G1 phase of the cell cycle. Growth arrested cells express the proto-oncogene c-myc and the gene for the CCAAT/enhancer binding protein (C/EBPα) between day 2 and 5. While c-Myc is strongly implicated in cell proliferation, C/EBPα is a differentiation-specific transcription factor with anti-proliferative activity. Here we have characterized the cell cycle arrest in differentiating 3T3-L1 cells. Arrested cells express the Cdk inhibitors p21 and p27, but, at the same time, show hyperphosphorylation of Rb and expression of the E2F-regulated thymidine kinase gene. The addition of new serum to arrested cells resulted in cyclin A expression and Cdk2 activity, but not in DNA synthesis. Simian virus 40 large tumor antigen (LTAg) is a potent mitogen. The mutant LTAg-K1, deficient in binding of pocket proteins and unable to induce DNA synthesis in serum-starved 3T3-L1 cells, efficiently induced DNA synthesis in differentiating 3T3-L1 cells. This indicates that pocket proteins are probably not involved in the control of the cell cycle arrest during 3T3-L1 cell differentiation. Our data suggest that the differentiation-specific cell cycle block in 3T3-L1 cells is resistant to high levels of c-Myc, inactivation of pocket proteins, upregulation of cyclin A levels, and Cdk2 activation, but can be abolished by a function of LTAg that is independent of binding to pocket proteins.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 4
Figure 5
Figure 3

Similar content being viewed by others

References

  • Birkenmeier EH, Gwynn B, Howard S, Jerry J, Gordon JI, Landschulz WH and McKnight SL. . 1989 Genes Dev. 3: 1146–1156.

  • Chen PL, Riley DJ, Chen Y and Lee WH. . 1996 Genes Dev. 10: 2794–2804.

  • Chirwing JM, Przybyla RJ, MacDonald RJ and Rutter WJ. . 1979 Biochemistry 18: 5294–5299.

    Article  CAS  Google Scholar 

  • Constance CM, Morgan J and Umek RM. . 1996 Mol. Cell. Biol. 16: 3878–3883.

    Article  CAS  Google Scholar 

  • Coppola JA and Cole MD. . 1986 Nature 320: 760–763.

    Article  CAS  Google Scholar 

  • Cornelius P, MacDougald OA and Lane MD. . 1994 Annu. Rev. Nutr. 14: 99–129.

    Article  CAS  Google Scholar 

  • Darlington G, Wang N and Hanson RW. . 1995 Curr. Opin. Gen. Dev. 5: 565–570.

  • Eilers M, Schirm S and Bishop JM. . 1991 EMBO J. 10: 133–141.

  • Fajas L, Fruchart JC and Auwerx J. . 1998 Curr. Opin. Cell Biol. 10: 165–171.

  • Fanning E and Knippers R. . 1992 Annu. Rev. Biochem. 61: 55–85.

    Article  CAS  Google Scholar 

  • Flores-Rozas H, Kelman Z, Dean FB, Pan ZQ, Harper JW, Elledge SJ, O'Donnell M and Hurwitz J. . 1994 Proc. Natl. Acad. Sci. USA 91: 8655–8659.

  • Freytag SO. . 1988 Mol. Cell. Biol. 8: 1614–1624.

    Article  CAS  Google Scholar 

  • Freytag SO and Geddes TJ. . 1992 Science 256: 379–382.

    Article  CAS  Google Scholar 

  • Gandarillas A and Watt FM. . 1997 Genes Dev. 11: 2869–2882.

  • Grana X and Reddy EP. . 1995 Oncogene 11: 211–219.

  • Gratzner HG, Ahmad PM, Stein J and Ahmad F. . (1985) Cytometry 6: 563–569.

    Article  CAS  Google Scholar 

  • Green H and Kehinde O. . 1975 Cell 5: 19–27.

    Article  CAS  Google Scholar 

  • Griep AE and Westphal H. . 1988 Proc. Natl. Acad. Sci. USA 85: 6806–6810.

  • Hermeking H and Eick D. . 1994 Science 265: 2091–2093.

    Article  CAS  Google Scholar 

  • Hermeking H, Wolf DA, Kohlhuber F, Dickmanns A, Billaud M, Fanning E and Eick D. . 1994 Proc. Natl. Acad. Sci. USA 91: 10412–10416.

  • Hoang AT, Cohen KJ, Barrett JF, Bergstrom DA and Dang CV. . 1994 Proc. Natl. Acad. Sci. USA 91: 6875–6879.

  • Hunter T and Pines J. . 1994 Cell 79: 573–582.

    Article  CAS  Google Scholar 

  • Jansen-Dürr P, Meichle A, Steiner P, Pagano M, Finke K, Botz J, Wessbecher J, Draetta G and Eilers M. . 1993 Proc. Natl. Acad. Sci. USA 90: 3685–3689.

  • Kalderon D and Smith AE. . 1984 Virology 139: 109–137.

    Article  CAS  Google Scholar 

  • Kempkes B, Spitkovsky D, Jansen-Dürr P, Ellwart JW, Kremmer E, Delecluse HJ, Rottenberger C, Bornkamm GW and Hammerschmidt W. . 1995 EMBO J. 14: 88–96.

  • Kiess M, Gill RM and Hamel PA. . 1995 Oncogene 10: 159–166.

  • Larsson LG, Ivhed I, Gidlund M, Pettersson U, Vennström B and Nilsson K. . 1988 Proc. Natl. Acad. Sci. USA 85: 2638–2642.

  • Li LH, Nerlov C, Prendergast G, MacGregor D and Ziff EB. . 1994a EMBO J. 13: 4070–4079.

  • Li R, Waga S, Hannon GJ, Beach D and Stillman B. . 1994b Nature 371: 534–537.

    Article  CAS  Google Scholar 

  • Lin FT and Lane MD. . 1994 Proc. Natl. Acad. Sci. USA 91: 8757–8761.

  • Lesage F, Hugnot JP, Amri EZ, Grimaldi P, Barhanin J and Lazdunski M. . 1994 Nucleic Acids Res. 22: 3685–3688.

  • MacArthur H and Walter G. . 1984 J. Virol. 52: 483–491.

  • MacDougald OA and Lane MD. . 1995 Annu. Rev. Biochem. 64: 345–373.

    Article  CAS  Google Scholar 

  • Meuth M and Green H. . 1974 Cell 3: 367–374.

    Article  CAS  Google Scholar 

  • Miner JH and Wold BJ. . 1991 Mol. Cell. Biol. 11: 2842–2851.

    Article  CAS  Google Scholar 

  • Morgan DO. . 1995 Nature 374: 131–134.

    Article  CAS  Google Scholar 

  • Müller R. . 1995 Trends Genetics 11: 173–178.

  • Müller D, Bouchard C, Rudolph B, Steiner P, Stuckmann I, Saffrich R, Ansorge W, Huttner W and Eilers M. . 1997 Oncogene 15: 2561–2576.

    Article  Google Scholar 

  • Ohkubo Y, Kishimoto T, Nakata T, Yasuda H and Endo T. . 1994 Exp. Cell Res. 214: 270–278.

  • Philipp A, Schneider A, Vasrik I, Finke K, Xiong Y, Beach D, Alitalo K and Eilers M. . 1994 Mol. Cell. Biol. 14: 4032–4043.

    Article  CAS  Google Scholar 

  • Prochownik EV and Kukowska J. . 1986 Nature 322: 848–850.

    Article  CAS  Google Scholar 

  • Rudolph B, Saffrich R, Zwicker J, Henglein B, Muller R, Ansorge W and Eilers M. . 1996 EMBO J. 15: 3065–3076.

  • Sambrook J, Fritsch EF and Maniatis T. . 1989 Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, ed. 2.

    Google Scholar 

  • Schneider MD, Perryman MB, Payne PA, Spizz G, Roberts R and Olson EN. . 1987 Mol. Cell. Biol. 7: 1973–1977.

    Article  CAS  Google Scholar 

  • Sherr JC. . 1995 Trends Biochem. Sci. 20: 187–190.

  • Tevosian SG, Shih HH, Mendelson KG, Sheppard KA, Paulson KE and Yee AS. . 1997 Genes Dev. 11: 383–396.

  • Timchenko NA, Wilde M, Nakanishi M, Smith JR and Darlington GJ. . 1996 Genes Dev. 10: 804–815.

  • Vlach J, Hennecke S, Alevizopoulos K, Conti D and Amati B. . 1996 EMBO J. 15: 6595–6604.

  • Waga S, Hannon GJ, Beach B and Stillman B. . 1994 Nature 369: 574–578.

    Article  CAS  Google Scholar 

  • Wolf DA, Hermeking H, Albert T, Herzinger T, Kind P and Eick D. . 1995 Oncogene 10: 2067–2078.

  • Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R and Beach D. . 1993 Nature 366: 701–704.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Drs C Sherr and P Jansen-Dürr for cyclin probes, W Kölch, C Karreman, F Kohlhuber, C Hall and T Müller for discussion and critical reading of the manuscript. Supported by the Deutsche Forschungsgemeinschaft (SFB190: Mechanismen und Faktoren der Genaktivierung), Deutsche Krebshilfe, European Union and Fonds der Chemischen Industrie.

Author information

Authors and Affiliations

  1. Institut für klinische Molekularbiologie und Tumorgenetik, GSF-Forschungszentrum für Umwelt und Gesundheit, Marchioninistrasse 25, 81377, München, Germany

    Manuela Reichert & Dirk Eick

Authors
  1. Manuela Reichert
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dirk Eick
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Reichert, M., Eick, D. Analysis of cell cycle arrest in adipocyte differentiation. Oncogene 18, 459–466 (1999). https://doi.org/10.1038/sj.onc.1202308

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1202308

Keywords

This article is cited by

Search

Advanced search

Quick links