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Canonical and atypical E2Fs regulate the mammalian endocycle

Nature Cell Biology volume 14, pages 1192–1202 (2012)Cite this article

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Abstract

The endocycle is a variant cell cycle consisting of successive DNA synthesis and gap phases that yield highly polyploid cells. Although essential for metazoan development, relatively little is known about its control or physiologic role in mammals. Using lineage-specific cre mice we identified two opposing arms of the E2F program, one driven by canonical transcription activation (E2F1, E2F2 and E2F3) and the other by atypical repression (E2F7 and E2F8), that converge on the regulation of endocycles in vivo. Ablation of canonical activators in the two endocycling tissues of mammals, trophoblast giant cells in the placenta and hepatocytes in the liver, augmented genome ploidy, whereas ablation of atypical repressors diminished ploidy. These two antagonistic arms coordinate the expression of a unique G2/M transcriptional program that is critical for mitosis, karyokinesis and cytokinesis. These results provide in vivo evidence for a direct role of E2F family members in regulating non-traditional cell cycles in mammals.

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Figure 1: Loss of E2f activators promotes trophoblast giant cells (TGC) and hepatocyte endocycles.
Figure 2: E2f7 and E2f8 promote TGC endocycles.
Figure 3: E2f8 is sufficient to promote hepatocyte endocycles.
Figure 4: Canonical activator and atypical repressor E2Fs regulate key transcriptional networks coordinating endocycles.
Figure 5: Atypical repressors E2F7/E2F8 directly bind gene targets involved in endocycle control.
Figure 6: Loss of E2f1 restores endocycles in E2f7/E2f8 deficient TGCs and hepatocytes.
Figure 7: Cyclin A ablation reinstates genome ploidy of E2f7/E2f8-deficient TGCs and hepatocytes.

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Acknowledgements

We thank L. Rawahneh, J. Moffitt and N. Lovett for excellent technical assistance with histology, and P. Wenzel for generating and collecting 123tko placentae. We also thank these individuals from OSUCCC Shared Resources: T. Wise, J. Palatini, H. Alders, P. Yan, P. Fada and B. Rodriguez (Microarray and Nucleic Acid Shared Resources); B. McElwain (Analytic Cytometry); R. Burry, K. Wolken, and B. Kemmenoe (Microscopy and Imaging); and K. La Perle (Comparative Pathology and Mouse Phenotyping). We are grateful for PL-1 antibodies provided by F. Talamantes (University of California, Santa Cruz, CA), Rcho-1 trophoblast stem cells from M. J. Soares (University of Kansas Medical Center, Kansas City, KS) and HepG2 cells from S. Jacobs. This work was funded by NIH grants to G.L. (R01CA85619, R01CA82259, R01HD047470, P01CA097189) and P.S. (R01CA132740). G.L. is a recipient of The Pew Charitable Trust Scholar Award and the Leukemia & Lymphoma Society Scholar Award. H-Z.C., M.M., S.S., S.R. and T. P. are recipients of the Pelotonia Fellowship Award.

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Author notes

  1. Jing Li

    Present address: Present address: Roche Molecular Systems, 4300 Hacienda Drive, Pleasanton, California 94588, USA,

  2. Hui-Zi Chen and Madhu M. Ouseph: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Molecular Virology, Department of Molecular Genetics, Solid Tumor Biology Program, Immunology and Medical Genetics, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, USA

    Hui-Zi Chen, Madhu M. Ouseph, Jing Li, Thierry Pécot, Veda Chokshi, Lindsey Kent, Sooin Bae, Morgan Byrne, Camille Duran, Grant Comstock, Prashant Trikha, Markus Mair, Shantibhusan Senapati, Chelsea K. Martin, Sagar Gandhi, Nicholas Wilson, Bin Liu, John C. Thompson, Tim Huang & Gustavo Leone

  2. Medical Scientist Training Program and Integrated Biomedical Graduate Program, College of Medicine, The Ohio State University, Columbus, Ohio 43210, USA

    Hui-Zi Chen & Gustavo Leone

  3. Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio 43210, USA

    Madhu M. Ouseph

  4. Computer Science and Engineering, The Ohio State University, Columbus, Ohio 43210, USA

    Thierry Pécot, Sundaresan Raman, Shantanu Singh, Marcelo Leone & Raghu Machiraju

  5. Biomedical Informatics, The Ohio State University, Columbus, Ohio 43210, USA

    Thierry Pécot, Bin Liu, Kun Huang & Victor Jin

  6. Department of Obstetrics and Gynecology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA

    Yi-Wen Huang

  7. Center for Biostatistics, Office of Health Sciences, The Ohio State University, Columbus, Ohio 43210, USA

    Xiaokui Mo & Soledad Fernandez

  8. Department of Cancer Biology, and Department of Genetics, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02215, USA

    Ilona Kalaszczynska & Piotr Sicinski

  9. Department of Genetics and Development, Department of Obstetrics and Gynecology, The Institute of Human Nutrition, and The Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, New York, New York 10032, USA

    Debra J. Wolgemuth

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Contributions

H-Z.C., M.M.O., J.L. and G.L. designed the experiments. H-Z.C., M.M.O., J.L., T.P., V.C., L.K. and S.B. performed experiments. H-Z.C. and M.M.O. co-wrote the paper with G.L.; all other authors listed helped perform experiments. Specifically, J.C.T., S.R., S.S., M.L., R.M. and K.H. helped with experiments relating to confocal microscopy and 3D reconstruction. X.M. and S.F. helped analyse data and performed statistics. I.K., D.J.W. and P.S. provided mice for the study and reviewed drafts of the manuscript. T.H., B.L. and V.J. contributed to discussion relating to Affymetrix/NanoString analyses.

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Correspondence to Gustavo Leone.

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Chen, HZ., Ouseph, M., Li, J. et al. Canonical and atypical E2Fs regulate the mammalian endocycle. Nat Cell Biol 14, 1192–1202 (2012). https://doi.org/10.1038/ncb2595

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