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.

Acute mutation of retinoblastoma gene function is sufficient for cell cycle re-entry

Abstract

Cancer cells arise from normal cells through the acquisition of a series of mutations in oncogenes and tumour suppressor genes1. Mouse models of human cancer often rely on germline alterations that activate or inactivate genes of interest. One limitation of this approach is that germline mutations might have effects other than somatic mutations, owing to developmental compensation2,3. To model sporadic cancers associated with inactivation of the retinoblastoma (RB) tumour suppressor gene in humans, we have produced a conditional allele of the mouse Rb gene. We show here that acute loss of Rb in primary quiescent cells is sufficient for cell cycle entry and has phenotypic consequences different from germline loss of Rb function. This difference is explained in part by functional compensation by the Rb-related gene p107. We also show that acute loss of Rb in senescent cells leads to reversal of the cellular senescence programme. Thus, the use of conditional knockout strategies might refine our understanding of gene function and help to model human cancer more accurately.

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: Acute deletion of Rb in MEFs.
Figure 2: Effects of acute versus germline loss of Rb function in quiescent cells.
Figure 3: Reversal of senescence after acute loss of Rb.

References

  1. 1

    Hanahan, D. & Weinberg, R. A. The hallmarks of cancer. Cell 100, 57–70 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2

    Van Dyke, T. & Jacks, T. Cancer modelling in the modern era: Progress and challenges. Cell 108, 135–144 (2002)

    CAS  Article  Google Scholar 

  3. 3

    Jonkers, J. & Berns, A. Conditional mouse models of sporadic cancer. Nature Rev. Cancer 2, 251–265 (2002)

    CAS  Article  Google Scholar 

  4. 4

    Sage, J. et al. Targeted disruption of the three Rb-related genes leads to loss of G1 control and immortalization. Genes Dev. 14, 3037–3050 (2000)

    CAS  Article  Google Scholar 

  5. 5

    MacPherson, D. et al. Conditional mutation of Rb causes cell cycle defects without apoptosis in the central nervous system. Mol. Cell. Biol. 23, 1044–1053 (2003)

    CAS  Article  Google Scholar 

  6. 6

    Hurford, R. K. Jr, Cobrinik, D., Lee, M. H. & Dyson, N. pRB and p107/p130 are required for the regulated expression of different sets of E2F responsive genes. Genes Dev. 11, 1447–1463 (1997)

    CAS  Article  Google Scholar 

  7. 7

    Lomazzi, M., Moroni, M. C., Jensen, M. R., Frittoli, E. & Helin, K. Suppression of the p53- or pRB-mediated G1 checkpoint is required for E2F-induced S-phase entry. Nature Genet. 31, 190–194 (2002)

    CAS  Article  Google Scholar 

  8. 8

    Trimarchi, J. M. & Lees, J. A. Sibling rivalry in the E2F family. Nature Rev. Mol. Cell Biol. 3, 11–20 (2002)

    CAS  Article  Google Scholar 

  9. 9

    Rubinson, D. A. et al. A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nature Genet. 33, 401–406 (2003)

    CAS  Article  Google Scholar 

  10. 10

    Hayflick, L. & Moorhead, P. S. The serial cultivation of human diploid cell strains. Exp. Cell Res. 25, 585–621 (1961)

    CAS  Article  Google Scholar 

  11. 11

    Campisi, J. Cellular senescence as a tumour-suppressor mechanism. Trends Cell Biol. 11, S27–S31 (2001)

    CAS  Article  Google Scholar 

  12. 12

    Schmitt, C. A. et al. A senescence program controlled by p53 and p16INK4a contributes to the outcome of cancer therapy. Cell 109, 335–346 (2002)

    CAS  Article  Google Scholar 

  13. 13

    Dannenberg, J. H., van Rossum, A., Schuijff, L. & te Riele, H. Ablation of the retinoblastoma gene family deregulates G1 control causing immortalization and increased cell turnover under growth-restricting conditions. Genes Dev. 14, 3051–3064 (2000)

    CAS  Article  Google Scholar 

  14. 14

    Carneiro, C. et al. p27 deficiency desensitizes Rb-/- cells to signals that trigger apoptosis during pituitary tumour development. Oncogene 22, 361–369 (2003)

    CAS  Article  Google Scholar 

  15. 15

    Classon, M. & Harlow, E. The retinoblastoma tumour suppressor in development and cancer. Nature Rev. Cancer 2, 910–917 (2002)

    CAS  Article  Google Scholar 

  16. 16

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

    CAS  Article  Google Scholar 

  17. 17

    Serrano, M., Lin, A. W., McCurrach, M. E., Beach, D. & Lowe, S. W. Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602 (1997)

    CAS  Article  Google Scholar 

  18. 18

    Jat, P. S. & Sharp, P. A. Cell lines established by a temperature-sensitive simian virus 40 large-T-antigen gene are growth restricted at the nonpermissive temperature. Mol. Cell. Biol. 9, 1672–1681 (1989)

    CAS  Article  Google Scholar 

  19. 19

    Gire, V. & Wynford-Thomas, D. Reinitiation of DNA synthesis and cell division in senescent human fibroblasts by microinjection of anti-p53 antibodies. Mol. Cell. Biol. 18, 1611–1621 (1998)

    CAS  Article  Google Scholar 

  20. 20

    Pfeifer, A., Brandon, E. P., Kootstra, N., Gage, F. H. & Verma, I. M. Delivery of the Cre recombinase by a self-deleting lentiviral vector: Efficient gene targeting in vivo. Proc. Natl Acad. Sci. USA 98, 11450–11455 (2001)

    CAS  Article  Google Scholar 

  21. 21

    Dirac, A. M. & Bernards, R. Reversal of senescence in mouse fibroblasts through lentiviral suppression of p53. J. Biol. Chem. 278, 11731–11734 (2003)

    CAS  Article  Google Scholar 

  22. 22

    Vooijs, M., van der Valk, M., te Riele, H. & Berns, A. Flp-mediated tissue-specific inactivation of the retinoblastoma tumour suppressor gene in the mouse. Oncogene 17, 1–12 (1998)

    CAS  Article  Google Scholar 

  23. 23

    Marino, S., Vooijs, M., van Der Gulden, H., Jonkers, J. & Berns, A. Induction of medulloblastomas in p53-null mutant mice by somatic inactivation of Rb in the external granular layer cells of the cerebellum. Genes Dev. 14, 994–1004 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  24. 24

    Brummelkamp, T. R., Bernards, R. & Agami, R. A system for stable expression of short interfering RNAs in mammalian cells. Science 296, 550–553 (2002)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank R. Jaenisch, S. Lowe and B. Kennedy for various useful reagents; D. Tuveson for helpful discussions; A. Brunet, K. Johnson and M. McLaughlin for critical reading of the manuscript; and members of F. Gertler's laboratory for help with the videomicroscopy experiments. This work was supported by funding from the Human Frontier Science Program, the Medical Foundation and the Merck/MIT postdoctoral fellowship program (J.S.), and from the National Cancer Institute and the Howard Hughes Medical Institute (T.J.).

Author information

Affiliations

Authors

Corresponding author

Correspondence to Tyler Jacks.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sage, J., Miller, A., Pérez-Mancera, P. et al. Acute mutation of retinoblastoma gene function is sufficient for cell cycle re-entry. Nature 424, 223–228 (2003). https://doi.org/10.1038/nature01764

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.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing