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DAP kinase activates a p19ARF/p53-mediated apoptotic checkpoint to suppress oncogenic transformation

Nature Cell Biology volume 3, pages 1–7 (2001)Cite this article

Abstract

DAP kinase is a pro-apoptotic calcium-regulated serine/threonine kinase, whose expression is frequently lost in human tumours. Here we show that DAP kinase counteracts oncogene-induced transformation by activating a p19ARF/p53-dependent apoptotic checkpoint. Ectopic expression of DAP kinase suppressed oncogenic transformation of primary embryonic fibroblasts by activating p53 in a p19ARF-dependent manner. Consequently, the fibroblasts underwent apoptosis, characterized by caspase activation and DNA fragmentation. In response to c-Myc or E2F-1, the endogenous DAP kinase protein was upregulated. Furthermore, functional or genetic inactivation of the endogenous DAP kinase reduced the extent of induction of p19ARF/p53 and weakened the subsequent apoptotic responses to c-Myc or E2F-1. These results establish a role for DAP kinase in an early apoptotic checkpoint designed to eliminate pre-malignant cells during cancer development.

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Figure 1: Suppression of transformation by DAP kinase in primary rodent fibroblasts.
Figure 2: Activation of p53 by DAP kinase.
Figure 3: DAP kinase induces p19ARF/p53-dependent apoptosis.
Figure 4: DAP-kinase-induced apoptosis requires the catalytic activity of the kinase, and involves caspase activation.
Figure 5: Inactivation of DAP kinase by a dominant-negative mutant reduces induction of p53 and apoptotic responses to c-Myc.
Figure 6: DAP-kinase-deficient MEFs display reduced extent of p53 induction by c-Myc or E2F-1.
Figure 7: E2F-1-induced apoptosis is attenuated in the absence of DAP kinase.

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Acknowledgements

We are grateful to Moshe Oren for helpful discussions and reagents. We thank Charles Sherr for the gift of p19ARF−/− MEFs, Claudio Schneider for kindly providing the anti-p21 antibodies, Tony Hunter for the membrane-targeted GFP plasmid, Doron Ginsberg for the E2F-1 plasmid, and members of the Kimchi lab for critical reading of the manuscript. This work was supported by the Israel Foundation, which is administered by the Israel Academy of Science and Humanities, and by QBI Ltd. M.S.H. is supported by the Forchheimer Foundation. R.A.D is an American Cancer Society Research Professor and is supported by grants from the National Institutes of Health. A.K. is the incumbent of the Helena Rubinstein Chair of Cancer Research.

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Authors and Affiliations

  1. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel

    Tal Raveh & Adi Kimchi

  2. Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, 10461 , USA

    Gustavo Droguett & Marshall S. Horwitz

  3. Departments of Medicine and Genetics, and Department of Adult Oncology, Harvard Medical School, Dana Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Ronald A. DePinho

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  1. Tal Raveh
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Correspondence to Adi Kimchi.

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Raveh, T., Droguett, G., Horwitz, M. et al. DAP kinase activates a p19ARF/p53-mediated apoptotic checkpoint to suppress oncogenic transformation. Nat Cell Biol 3, 1–7 (2001). https://doi.org/10.1038/35050500

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