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E2F8 is essential for polyploidization in mammalian cells

Nature Cell Biology volume 14, pages 1181–1191 (2012)Cite this article

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Abstract

Polyploidization is observed in all mammalian species and is a characteristic feature of hepatocytes, but its molecular mechanism and biological significance are unknown. Hepatocyte polyploidization in rodents occurs through incomplete cytokinesis, starts after weaning and increases with age. Here, we show in mice that atypical E2F8 is induced after weaning and required for hepatocyte binucleation and polyploidization. A deficiency in E2f8 led to an increase in the expression level of E2F target genes promoting cytokinesis and thereby preventing polyploidization. In contrast, loss of E2f1 enhanced polyploidization and suppressed the polyploidization defect of hepatocytes deficient for atypical E2Fs. In addition, E2F8 and E2F1 were found on the same subset of target promoters. Contrary to the long-standing hypothesis that polyploidization indicates terminal differentiation and senescence, we show that prevention of polyploidization through inactivation of atypical E2Fs has, surprisingly, no impact on liver differentiation, zonation, metabolism and regeneration. Together, these results identify E2F8 as a repressor and E2F1 as an activator of a transcriptional network controlling polyploidization in mammalian cells.

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Figure 1: Atypical E2Fs are induced during the onset of hepatocyte polyploidization.
Figure 2: E2F8 is essential for hepatocyte binucleation.
Figure 3: E2F8 is essential for hepatocyte polyploidization.
Figure 4: Inactivation of atypical E2Fs increases E2F target gene expression.
Figure 5: Inactivation of atypical E2Fs increases cytokinetic gene expression.
Figure 6: E2F8 and E2F1 bind to the same promoters of cytokinetic genes.
Figure 7: Loss of E2f1 restores polyploidization in E2f7−/−E2f8−/− livers.
Figure 8: Polyploidization has no impact on hepatic differentiation or regeneration.

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Acknowledgements

We thank R. Medema, R. Klompmaker (both at the Department of Medical Oncology, University Medical Center Utrecht, Utrecht, and The Netherlands Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands), E. Cuppen (Hubrecht Institute, University Medical Center Utrecht, Cancer Genomics Center, Utrecht, The Netherlands), G. Leone (Department of Molecular Virology, Immunology and Medical Genetics, Comprehensive Cancer Center, College of Medicine, The Ohio State University, Columbus, USA), V. Guyrev, J. Mul (both at Hubrecht Institute, as above) and J. Mol (Department of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands) for reagents, mice, technical assistance, bioinformatic support, and advice, and W. Bakker and B. Weijts for critical comments on the manuscript.

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  1. Shusil K. Pandit and Bart Westendorp: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, 3584CL Utrecht, The Netherlands

    Shusil K. Pandit, Bart Westendorp, Sathidpak Nantasanti, Elsbeth van Liere, Peter C. J. Tooten, Peter W. A. Cornelissen, Mathilda J. M. Toussaint & Alain de Bruin

  2. University of Amsterdam, Academic Medical Center, Tytgat Institute for Liver and Intestinal Research, 1105BK Amsterdam, The Netherlands

    Wouter H. Lamers

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Contributions

S.K.P. carried out all in vivo experiments assisted by S.N., P.C.J.T. and M.J.M.T. B.W. performed the microarray analysis and the ChIP assays. E.v.L. and P.W.A.C. performed the qPCR analysis. W.H.L. performed the zonation analysis. S.K.P., B.W. and A.d.B designed the experiments and wrote the paper. All authors discussed the results and edited the manuscript.

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Correspondence to Alain de Bruin.

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Pandit, S., Westendorp, B., Nantasanti, S. et al. E2F8 is essential for polyploidization in mammalian cells. Nat Cell Biol 14, 1181–1191 (2012). https://doi.org/10.1038/ncb2585

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