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Phosphorylation of Dok1 by Abl family kinases inhibits CrkI transforming activity

Oncogene volume 34, pages 2650–2659 (2015)Cite this article

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Abstract

The Crk SH2/SH3 adaptor and the Abl nonreceptor tyrosine kinase were first identified as oncoproteins, and both can induce tumorigenesis when overexpressed or mutationally activated. We previously reported the surprising finding that inhibition or knockdown of Abl family kinases enhanced transformation of mouse fibroblasts by CrkI. Abl family inhibitors are currently used or are being tested for treatment of human malignancies, and our finding raised concerns that such inhibitors might actually promote the growth of tumors overexpressing CrkI. Here, we identify the Dok1 adaptor as the key effector for the enhancement of CrkI transformation by Abl inhibition. We show that phosphorylation of tyrosines 295 and 361 of Dok1 by Abl family kinases suppresses CrkI transforming activity, and that upon phosphorylation these tyrosines bind the SH2 domains of the Ras inhibitor p120 RasGAP. Knockdown of RasGAP resulted in a similar enhancement of CrkI transformation, consistent with a critical role for Ras activity. Imaging studies using a FRET sensor of Ras activation revealed alterations in the localization of activated Ras in CrkI-transformed cells. Our results support a model in which Dok1 phosphorylation normally suppresses localized Ras pathway activity in Crk-transformed cells via recruitment and/or activation of RasGAP, and that preventing this negative feedback mechanism by inhibiting Abl family kinases leads to enhanced transformation by Crk.

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Acknowledgements

We thank Sofya Borinskaya for technical help in soft agar colony counting, Mari Ogiue-Ikeda for the preparation of the SH2 probes and Joshua Jadwin for critically reading this manuscript. In addition, we greatly appreciate Pier Paolo Pandolfi from Harvard Medical School for generously providing the human Dok1 cDNA. This study was supported by grants CA82258 (to BJM) and NS071216 (to YIW) from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Genetics and Developmental Biology, Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, University of Connecticut Health Center, Farmington, CT, USA,

    K Y Ng, K Machida & B J Mayer

  2. Department of Genetics and Developmental Biology, Richard D. Berlin Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT, USA

    T Yin, Y I Wu & B J Mayer

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Correspondence to B J Mayer.

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Ng, K., Yin, T., Machida, K. et al. Phosphorylation of Dok1 by Abl family kinases inhibits CrkI transforming activity. Oncogene 34, 2650–2659 (2015). https://doi.org/10.1038/onc.2014.210

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