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 Article
  • Published:

Targeting an E2F site in the mouse genome prevents promoter silencing in quiescent and post-mitotic cells

Abstract

Previous studies have shown that the cell cycle-regulated B-myb promoter contains a conserved E2F binding site that is critical for repressing transcription in quiescent cells. To investigate its significance for permanent promoter silencing, we have inactivated this binding site in the mouse genome. Mice homozygous for the mutant B-mybmE2F allele were fully viable, however, B-myb transcription was derepressed during quiescence in mouse embryo fibroblasts (MEFs) derived from mutant animals. Moreover, it was found that mutation of the E2F site resulted in abnormal maintenance of B-myb expression in senescent MEFs and in differentiated brain tissue. These findings therefore reveal a direct and primary role for repressive E2F complexes in silencing gene expression in post-mitotic cells. Analysis of histone modifications at the promoter showed that histone H3 lysine 9 was constitutively acetylated throughout the cell cycle in homozygous mutant MEFs. This mouse system is the first description of an E2F site mutation in situ and will facilitate the study of E2F function in vivo.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  • Attwooll C, Lazzerini Denchi E, Helin K . (2004). The E2F family: specific functions and overlapping interests. EMBO J 23: 4709–4716.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bennett JD, Farlie P, Watson RJ . (1996). E2F binding is required but not sufficient for repression of B-Myb transcription in quiescent fibroblasts. Oncogene 13: 1073–1082.

    CAS  PubMed  Google Scholar 

  • Catchpole S, Tavner F, Le Cam L, Sardet C, Watson RJ . (2002). A B-myb promoter corepressor site facilitates in vivo occupation of the adjacent E2F site by p107 × E2F and p130 × E2F complexes. J Biol Chem 277: 39015–39024.

    Article  CAS  PubMed  Google Scholar 

  • Cobrinik D . (2005). Pocket proteins and cell cycle control. Oncogene 24: 2796–2809.

    Article  CAS  PubMed  Google Scholar 

  • Gaubatz S, Lindeman GJ, Ishida S, Jakoi L, Nevins JR, Livingston DM et al. (2000). E2F4 and E2F5 play an essential role in pocket protein-mediated G1 control. Mol Cell 6: 729–735.

    Article  CAS  PubMed  Google Scholar 

  • Hurford RK, Cobrinik D, Lee M-H, Dyson N . (1997). pRb and p107/p130 are required for the regulated expression of different sets of E2F responsive genes. Genes Dev 11: 1447–1463.

    Article  CAS  PubMed  Google Scholar 

  • Joaquin M, Watson RJ . (2003). Cell cycle regulation by the B-Myb transcription factor. Cell Mol Life Sci 60: 2389–2401.

    Article  CAS  PubMed  Google Scholar 

  • Lam EW, Robinson C, Watson RJ . (1992). Characterization and cell cycle-regulated expression of mouse B-myb. Oncogene 7: 1885–1890.

    CAS  PubMed  Google Scholar 

  • Lam EW-F, Watson RJ . (1993). An E2F-binding site mediates cell-cycle regulated repression of mouse B-myb transcription. EMBO J 12: 2705–2713.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lane S, Farlie P, Watson R . (1997). B-Myb function can be markedly enhanced by cyclin A-dependent kinase and protein truncation. Oncogene 14: 2445–2453.

    Article  CAS  PubMed  Google Scholar 

  • Liu DX, Greene LA . (2001). Regulation of neuronal survival and death by E2F-dependent gene repression and derepression. Neuron 32: 425–438.

    Article  CAS  PubMed  Google Scholar 

  • Maehara K, Yamakoshi K, Ohtani N, Kubo Y, Takahashi A, Arase S et al. (2005). Reduction of total E2F/DP activity induces senescence-like cell cycle arrest in cancer cells lacking functional pRB and p53. J Cell Biol 168: 553–560.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Margueron R, Trojer P, Reinberg D . (2005). The key to development: interpreting the histone code? Curr Opin Genet Dev 15: 163–176.

    Article  CAS  PubMed  Google Scholar 

  • Morgenstern JP, Land H . (1990). Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res 18: 3587–3596.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nagy A, Gertsenstein M, Vintersten K, Behringer R . (2003). Manipulating the Mouse Genome; a Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York.

    Google Scholar 

  • Narita M, Nunez S, Heard E, Lin AW, Hearn SA, Spector D et al. (2003). Rb-mediated heterochromatin formation and silencing of E2F target genes during cellular senescence. Cell 113: 703–716.

    Article  CAS  PubMed  Google Scholar 

  • Nielsen SJ, Schneider R, Bauer UM, Bannister AJ, Morrison A, O'Carroll D et al. (2001). Rb targets histone H3 methylation and HP1 to promoters. Nature 412: 561–565.

    Article  CAS  PubMed  Google Scholar 

  • Rayman JB, Takahashi Y, Dannenberg JH, Catchpole S, Watson R, te Riele H et al. (2002). E2F mediates cell cycle-dependent transcriptional repression in vivo by recruitment of specific co-repressor complex. Genes Dev 16: 933–947.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Robinson C, Light Y, Groves R, Mann D, Marais R, Watson R . (1996). Cell-cycle regulation of B-Myb protein expression: specific phosphorylation during the S phase of the cell cycle. Oncogene 12: 1855–1864.

    CAS  PubMed  Google Scholar 

  • Rowland BD, Denissov SG, Douma S, Stunnenberg HG, Bernards R, Peeper DS . (2002). E2F transcriptional repressor complexes are critical downstream targets of p19(ARF)/p53-induced proliferative arrest. Cancer Cell 2: 55–65.

    Article  CAS  PubMed  Google Scholar 

  • Sala A, Kundu M, Casella I, Engelhard A, Calabretta B, Grasso L et al. (1997). Activation of human B-MYB by cyclins. Proc Natl Acad Sci USA 94: 532–536.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sitzmann J, Noben-Trauth K, Kamano H, Klempnauer K-H . (1996). Expression of B-Myb during mouse embryogenesis. Oncogene 12: 1889–1894.

    CAS  PubMed  Google Scholar 

  • Takahashi Y, Rayman JB, Dynlacht BD . (2000). Analysis of promoter binding by the E2F and pRB families in vivo: distinct E2F proteins mediate activation and repression. Genes Dev 14: 804–816.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Todaro GJ, Green H . (1963). Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol 17: 299–313.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wells J, Boyd KE, Fry CJ, Bartley SM, Farnham PJ . (2000). Target gene specificity of E2F and pocket protein family members in living cells. Mol Cell Biol 20: 5797–5807.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ziebold U, Bartsch O, Marais R, Ferrari S, Klempnauer K-H . (1997). Phosphorylation and activation of B-Myb by cyclin A-Cdk2. Curr Biol 7: 253–260.

    Article  CAS  PubMed  Google Scholar 

  • Zwicker J, Liu N, Engeland K, Lucibello FC, Müller R . (1996). Cell cycle regulation of E2F site occupation in vivo. Science 271: 1595–1597.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Paloma Garcia, Nikla Emambokus, Gordon Peters, Xin Lu and Laura O'Neill for reagents and valuable advice. This work was supported by project grants from Association for International Cancer Research (to RJW and JF) and Biotechnology and Biological Sciences Research Council (to RJW).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R J Watson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tavner, F., Frampton, J. & Watson, R. Targeting an E2F site in the mouse genome prevents promoter silencing in quiescent and post-mitotic cells. Oncogene 26, 2727–2735 (2007). https://doi.org/10.1038/sj.onc.1210087

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210087

Keywords

This article is cited by

Search

Quick links