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.
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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.
The authors declare no competing financial interests.
This file contains Supplementary Figures 1-20 with Legends. (PDF 4152 kb)
Supplementary Table 1 shows gene expression changed (villi vs crypts). (XLS 3077 kb)
Supplementary Table shows gene expression changed in RbKO villi and crypts. (XLS 1895 kb)
This table shows gene ontology RbKO villi and crypts. (XLS 419 kb)
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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
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