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E2F4 regulates a stable G2 arrest response to genotoxic stress in prostate carcinoma

Oncogene volume 26, pages 1897–1909 (2007)Cite this article

Abstract

The retinoblastoma (pRB) family proteins regulate the E2F transcription factors; their complexes regulate critical transitions through the cell cycle. The function of these pRB family/E2F complexes, which includes p130/E2F4, in response to genotoxic agents, is not well understood. We investigated the role of E2F4 in the genotoxic stress response. Following radiation treatment, E2F4 colocalized with p130 in the nucleus during a radiation-induced stable G2-phase arrest. Arrested cells had significantly decreased expression of Cyclins A2 and B1 and decreased phosphorylation of mitotic protein monoclonal-2 (MPM-2) mitotic proteins. Small interference RNA (siRNA)-mediated knockdown of E2F4 sensitized cells to subsequent irradiation, resulting in enhanced cellular DNA damage and cell death, as determined by caspase activation and decreased clonogenic cell survival. Downstream E2F4 targets potentially involved in the progression from G2 into M phase were identified by oligonucleotide microarray expression profiling. Chromatin immunoprecipitation localized E2F4 at promoter regions of the Bub3 and Pttg1 mitotic genes following irradiation, which were among the downregulated genes identified by the microarray. These data suggest that in response to radiation, E2F4 becomes active in the nucleus, enforces a stable G2 arrest by target gene repression, and thus provides increased cell survival ability by minimizing propagation of cells that have irreparable DNA damage.

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Acknowledgements

We thank Drs W Heston and A Gudkov (Cleveland Clinic) for valuable reagents and B Dynlacht (NYU) for advice. We also thank the Flow Cytometry and Imaging Core facilities staff at the Cleveland Clinic Foundation, and RM Sramkowski at the Case Comprehensive Cancer Center for their invaluable help. This work was supported by research grants from the US National Institute of Health (CA81504 and CA82858).

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Authors and Affiliations

  1. Department of Cancer Biology, Lerner Research Institute, Cleveland, OH, USA

    M E Crosby, D Gupta & A Almasan

  2. Department of Environmental Health Sciences, Case Western Reserve University, Cleveland, OH, USA

    M E Crosby & A Almasan

  3. Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, USA

    J Jacobberger

  4. Department of Radiation Oncology, Cleveland Clinic, Cleveland, OH, USA

    R M Macklis & A Almasan

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  1. M E Crosby
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Correspondence to A Almasan.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Crosby, M., Jacobberger, J., Gupta, D. et al. E2F4 regulates a stable G2 arrest response to genotoxic stress in prostate carcinoma. Oncogene 26, 1897–1909 (2007). https://doi.org/10.1038/sj.onc.1209998

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