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Accelerated DNA replication in E2F1- and E2F2-deficient macrophages leads to induction of the DNA damage response and p21CIP1-dependent senescence

Oncogene volume 29, pages 5579–5590 (2010)Cite this article

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Abstract

E2F1–3 proteins appear to have distinct roles in progenitor cells and in differentiating cells undergoing cell cycle exit. However, the function of these proteins in paradigms of terminal differentiation that involve continued cell division has not been examined. Using compound E2F1/E2F2-deficient mice, we have examined the effects of E2F1 and E2F2 loss on the differentiation and simultaneous proliferation of bone-marrow-derived cells toward the macrophage lineage. We show that E2F1/E2F2 deficiency results in accelerated DNA replication and cellular division during the initial cell division cycles of bone-marrow-derived cells, arguing that E2F1/E2F2 are required to restrain proliferation of pro-monocyte progenitors during their differentiation into macrophages, without promoting their cell cycle exit. Accelerated proliferation is accompanied by early expression of DNA replication and cell cycle regulators. Remarkably, rapid proliferation of E2F1/E2F2 compound mutant cultures is temporally followed by induction of a DNA damage response and the implementation of a p21CIP1-dependent senescence. We further show that differentiating E2F1/E2F2-knockout macrophages do not trigger a DNA damage response pathway in the absence of DNA replication. These findings underscore the relevance of E2F1 and E2F2 as suppressors of hematopoietic progenitor expansion. Our data indicate that their absence in differentiating macrophages initiates a senescence program that results from enforcement of a DNA damage response triggered by DNA hyper-replication.

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Acknowledgements

We thank members of the Zubiaga laboratory for helpful discussions, Naiara Zorrilla for technical support and Dimitri Balomenos for p21CIP1−/− mice. AI is a recipient of the University of the Basque Country postdoctoral fellowship. OZ and JF are recipients of Basque Government fellowships for graduate students. This work was supported by grants to AMZ from the Spanish Ministry of Science and Innovation (SAF2009-12037 and Consolider-Ingenio 2010 Programme, CSD2007-00017) and the Basque Government Department of Industry (Etortek-IE06-178).

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Authors and Affiliations

  1. Department of Genetics, Physical Anthropology and Animal Physiology, University of the Basque Country, Leioa, Bilbao, Spain

    A Iglesias-Ara, O Zenarruzabeitia, J Fernandez-Rueda & A M Zubiaga

  2. Macrophage Biology Group, Institute for Research in Biomedicine (IRB Barcelona) and University of Barcelona, Barcelona, Spain

    E Sánchez-Tilló & A Celada

  3. Division of Endocrinology and Metabolism, University of California, San Diego, La Jolla, CA, USA

    S J Field

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  1. A Iglesias-Ara
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Correspondence to A M Zubiaga.

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Iglesias-Ara, A., Zenarruzabeitia, O., Fernandez-Rueda, J. et al. Accelerated DNA replication in E2F1- and E2F2-deficient macrophages leads to induction of the DNA damage response and p21CIP1-dependent senescence. Oncogene 29, 5579–5590 (2010). https://doi.org/10.1038/onc.2010.296

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