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DAPK2 is a novel E2F1/KLF6 target gene involved in their proapoptotic function

Oncogene volume 27, pages 5706–5716 (2008)Cite this article

Abstract

Death-associated protein kinase 2 (DAPK2) belongs to a family of proapoptotic Ca2+/calmodulin-regulated serine/threonine kinases. We recently identified DAPK2 as an enhancing factor during granulocytic differentiation. To identify transcriptional DAPK2 regulators, we cloned 2.7 kb of the 5′-flanking region of the DAPK2 gene. We found that E2F1 and Krüppel-like factor 6 (KLF6) strongly activate the DAPK2 promoter. We mapped the E2F1 and KLF6 responsive elements to a GC-rich region 5′ of exon 1 containing several binding sites for KLF6 and Sp1 but not for E2F. Moreover, we showed that transcriptional activation of DAPK2 by E2F1 and KLF6 is dependent on Sp1 using Sp1/KLF6-deficient insect cells, mithramycin A treatment to block Sp1-binding or Sp1 knockdown cells. Chromatin immunoprecipitation revealed recruitment of Sp1 and to lesser extent that of E2F1 and KLF6 to the DAPK2 promoter. Activation of E2F1 in osteosarcoma cells led to an increase of endogenous DAPK2 paralleled by cell death. Inhibition of DAPK2 expression resulted in significantly reduced cell death upon E2F1 activation. Similarly, KLF6 expression in H1299 cells increased DAPK2 levels accompanied by cell death that is markedly decreased upon DAPK2 knockdown. Moreover, E2F1 and KLF6 show cooperation in activating the DAPK2 promoter. In summary, our findings establish DAPK2 as a novel Sp1-dependent target gene for E2F1 and KLF6 in cell death response.

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Acknowledgements

Expert technical assistance by D Shan and G Arvidsson is appreciated. We are grateful to Drs SL Friedmann, G Thiel and JJ Rossi for providing the KLF6, CBP/p300 and Sp1 expression plasmids as well as an U6 promoter constructs. We also thank Dr D Kojic for providing the Schneider SL2 cells. This work was supported by grants from the Swiss National Foundation 3100-067213 (to AT and MFF); the Marlies-Schwegler Foundation and the Bernese Foundation of Cancer Research (to MFF and AT); the Werner and Hedy Berger-Janser Foundation of Cancer Research (to MFF and MPT); the Bern University Research Foundation (to MPT); and the Danish Cancer Society (to KH).

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Authors and Affiliations

  1. Department of Clinical Research, Experimental Oncology/Hematology, University of Bern, Bern, Switzerland

    A Britschgi, M Rizzi, M Jenal, A Ress, A Tobler, M F Fey & M P Tschan

  2. Centre for Epigenetics, Biotech Research and Innovation Centre, University of Copenhagen, Copenhagen, Denmark

    E Trinh & K Helin

  3. Hematology Department, Inselspital, Bern University Hospital, Bern, Switzerland

    A Tobler

  4. Medical Oncology Department, Inselspital, Bern University Hospital, Bern, Switzerland

    M F Fey

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  1. A Britschgi
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Correspondence to M P Tschan.

Additional information

Authors contributions: AB performed the experimental research, interpreted the data and drafted the article. ET performed and analysed E2F1 activation and ChIP experiments, MR cloned promoter and lentiviral constructs, performed reporter assays. MJ performed and analysed U2OS expression experiments. AR performed immunoprecipitations. AT, MFF and KH instigated the initial concept and experimental design and revised the drafted article. MPT designed the project, analysed data and gave final approval of the submitted paper.

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

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Britschgi, A., Trinh, E., Rizzi, M. et al. DAPK2 is a novel E2F1/KLF6 target gene involved in their proapoptotic function. Oncogene 27, 5706–5716 (2008). https://doi.org/10.1038/onc.2008.179

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