E2f1–3 switch from activators in progenitor cells to repressors in differentiating cells

Abstract

In the established model of mammalian cell cycle control, the retinoblastoma protein (Rb) functions to restrict cells from entering S phase by binding and sequestering E2f activators (E2f1, E2f2 and E2f3), which are invariably portrayed as the ultimate effectors of a transcriptional program that commit cells to enter and progress through S phase1,2. Using a panel of tissue-specific cre-transgenic mice and conditional E2f alleles we examined the effects of E2f1, E2f2 and E2f3 triple deficiency in murine embryonic stem cells, embryos and small intestines. We show that in normal dividing progenitor cells E2f1–3 function as transcriptional activators, but contrary to the current view, are dispensable for cell division and instead are necessary for cell survival. In differentiating cells E2f1–3 function in a complex with Rb as repressors to silence E2f targets and facilitate exit from the cell cycle. The inactivation of Rb in differentiating cells resulted in a switch of E2f1–3 from repressors to activators, leading to the superactivation of E2f responsive targets and ectopic cell divisions. Loss of E2f1–3 completely suppressed these phenotypes caused by Rb deficiency. This work contextualizes the activator versus repressor functions of E2f1–3 in vivo, revealing distinct roles in dividing versus differentiating cells and in normal versus cancer-like cell cycles.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Cell proliferation in the absence of E2f1–3.
Figure 2: Apoptosis of crypt intestinal cells in the absence of E2f1, E2f2 and E2f3.
Figure 3: Repression of E2f target genes in E2f1–3 -deficient villi.
Figure 4: E2f1–3 contribute to the ectopic cell proliferation caused by Rb -deficiency.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

All microarray data have been deposited at the Gene Expression Omnibus at the National Center for Biotechnology Information under accession number GSE16454.

References

  1. 1

    Iaquinta, P. J. & Lees, J. A. Life and death decisions by the E2F transcription factors. Curr. Opin. Cell Biol. 19, 649–657 (2007)

  2. 2

    Dimova, D. K. & Dyson, N. J. The E2F transcriptional network: old acquaintances with new faces. Oncogene 24, 2810–2826 (2005)

  3. 3

    DeGregori, J., Leone, G., Miron, A., Jakoi, L. & Nevins, J. R. Distinct roles for E2F proteins in cell growth control and apoptosis. Proc. Natl Acad. Sci. USA 94, 7245–7250 (1997)

  4. 4

    Johnson, D. G., Schwarz, J. K., Cress, W. D. & Nevins, J. R. Expression of transcription factor E2F1 induces quiescent cells to enter S phase. Nature 365, 349–352 (1993)

  5. 5

    Wu, L. et al. The E2F1–3 transcription factors are essential for cellular proliferation. Nature 414, 457–462 (2001)

  6. 6

    Timmers, C. et al. E2f1, E2f2, and E2f3 control E2F target expression and cellular proliferation via a p53-dependent negative feedback loop. Mol. Cell. Biol. 27, 65–78 (2007)

  7. 7

    Sharma, N. et al. Control of the p53-p21CIP1 axis by E2f1, E2f2, and E2f3 is essential for G1/S progression and cellular transformation. J. Biol. Chem. 281, 36124–36131 (2006)

  8. 8

    Saenz-Robles, M. T. et al. Intestinal hyperplasia induced by simian virus 40 large tumor antigen requires E2F2. J. Virol. 81, 13191–13199 (2007)

  9. 9

    Rowland, B. D. & Bernards, R. Re-evaluating cell-cycle regulation by E2Fs. Cell 127, 871–874 (2006)

  10. 10

    Murga, M. et al. Mutation of E2F2 in mice causes enhanced T lymphocyte proliferation, leading to the development of autoimmunity. Immunity 15, 959–970 (2001)

  11. 11

    Iglesias, A. et al. Diabetes and exocrine pancreatic insufficiency in E2F1/E2F2 double-mutant mice. J. Clin. Invest. 113, 1398–1407 (2004)

  12. 12

    Cloud, J. E. et al. Mutant mouse models reveal the relative roles of E2F1 and E2F3 in vivo . Mol. Cell. Biol. 22, 2663–2672 (2002)

  13. 13

    van der Flier, L. G. & Clevers, H. Stem cells, self-renewal, and differentiation in the intestinal epithelium. Annu. Rev. Physiol. 71, 241–260 (2009)

  14. 14

    Ireland, H. et al. Inducible Cre-mediated control of gene expression in the murine gastrointestinal tract: effect of loss of β-catenin. Gastroenterology 126, 1236–1246 (2004)

  15. 15

    Chen, D. et al. Division and apoptosis of E2f-deficient retinal progenitors. Nature 10.1038/nature08544 (this issue)

  16. 16

    Coopersmith, C. M. & Gordon, J. I. γ-Ray-induced apoptosis in transgenic mice with proliferative abnormalities in their intestinal epithelium: re-entry of villus enterocytes into the cell cycle does not affect their radioresistance but enhances the radiosensitivity of the crypt by inducing p53. Oncogene 15, 131–141 (1997)

  17. 17

    Mootha, V. K. et al. PGC-1α-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nature Genet. 34, 267–273 (2003)

  18. 18

    Kong, L. J., Chang, J. T., Bild, A. H. & Nevins, J. R. Compensation and specificity of function within the E2F family. Oncogene 26, 321–327 (2007)

  19. 19

    Xu, X. et al. A comprehensive ChIP-chip analysis of E2F1, E2F4, and E2F6 in normal and tumor cells reveals interchangeable roles of E2F family members. Genome Res. 17, 1550–1561 (2007)

  20. 20

    Chong, J. L. et al. E2f3a and E2f3b contribute to the control of cell proliferation and mouse development. Mol. Cell. Biol. 29, 414–424 (2009)

  21. 21

    Leone, G. et al. E2F3 activity is regulated during the cell cycle and is required for the induction of S phase. Genes Dev. 12, 2120–2130 (1998)

  22. 22

    Leone, G. et al. Identification of a novel E2F3 product suggests a mechanism for determining specificity of repression by Rb proteins. Mol. Cell. Biol. 20, 3626–3632 (2000)

  23. 23

    Eisen, M. B., Spellman, P. T., Brown, P. O. & Botstein, D. Cluster analysis and display of genome-wide expression patterns. Proc. Natl Acad. Sci. USA 95, 14863–14868 (1998)

  24. 24

    Malumbres, M. & Barbacid, M. Mammalian cyclin-dependent kinases. Trends Biochem. Sci. 30, 630–641 (2005)

  25. 25

    Martín, A. et al. Cdk2 is dispensable for cell cycle inhibition and tumor suppression mediated by p27Kip1 and p21Cip1 . Cancer Cell 7, 591–598 (2005)

  26. 26

    Malumbres, M. et al. Mammalian cells cycle without the D-type cyclin-dependent kinases Cdk4 and Cdk6. Cell 118, 493–504 (2004)

  27. 27

    Russell, P. & Nurse, P. Schizosaccharomyces pombe and Saccharomyces cerevisiae: a look at yeasts divided. Cell 45, 781–782 (1986)

  28. 28

    Helin, K. et al. A cDNA encoding a pRB-binding protein with properties of the transcription factor E2F. Cell 70, 337–350 (1992)

  29. 29

    Kaelin, W. G. et al. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. Cell 70, 351–364 (1992)

  30. 30

    Nevins, J. R. Transcriptional regulation. A closer look at E2F. Nature 358, 375–376 (1992)

  31. 31

    Haigis, K., Sage, J., Glickman, J., Shafer, S. & Jacks, T. The related retinoblastoma (pRb) and p130 proteins cooperate to regulate homeostasis in the intestinal epithelium. J. Biol. Chem. 281, 638–647 (2006)

Download references

Acknowledgements

We thank L. Rawahneh, J. Moffitt and R. Rajmohan for technical assistance with histology. We also thank A. de Bruin and S. Naidu for assistance in analysing histological slides. We are thankful to J. Groden, A. Simcox and D. Guttridge for their critical comments. This work was funded by NIH grants to G.L. (R01CA85619, R01CA82259, R01HD04470, P01CA097189) and NIH grant to J.M.P. (CA098956); J.-L.C. is the recipient of a DoD award (BC061730). P.L.W. was supported by NIH training grant 5 T32 CA106196-04.

Author Contributions M.L.R., J.M.P. and G.L. designed and supervised this study, analysed data, and helped write and edit the manuscript. J.-L.C., P.L.W. and M.T.S.-R. designed and performed experiments, collected and analysed data, and co-wrote the paper. V.N., A.F., Y.M.G., N.S., H.-Z.C., M.O., S.-H.W., P.T., B.C. and L.M. technically assisted with experiments and collected and analysed data. D.C. and R.B. performed and analysed gene expression of retina. J.P.H. and P.G.C. contributed to the analysis and comparison of gene microarray data. D.J.W. and O.J.S. contributed to the generation of key reagents.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Gustavo Leone.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-20 with Legends. (PDF 4152 kb)

Supplementary Table 1

Supplementary Table 1 shows gene expression changed (villi vs crypts). (XLS 3077 kb)

Supplementary Table 2

Supplementary Table shows gene expression changed in RbKO villi and crypts. (XLS 1895 kb)

Supplementary Table 3

This table shows gene ontology RbKO villi and crypts. (XLS 419 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Chong, J., Wenzel, P., Sáenz-Robles, M. et al. E2f1–3 switch from activators in progenitor cells to repressors in differentiating cells. Nature 462, 930–934 (2009). https://doi.org/10.1038/nature08677

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.