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Diacylglycerol kinase-α phosphorylation by Src on Y335 is required for activation, membrane recruitment and Hgf-induced cell motility

Oncogene volume 27, pages 942–956 (2008)Cite this article

Abstract

Diacylglycerol (DAG) kinases (Dgk), which phosphorylate DAG to generate phosphatidic acid, act as either positive or negative key regulators of cell signaling. We previously showed that Src mediates growth factors-induced activation of Dgk-α, whose activity is required for cell motility, proliferation and angiogenesis. Here, we demonstrate that both hepatocytes growth factor (HGF) stimulation and v-Src transformation induce tyrosine phosphorylation of Dgk-α on Y335, through a mechanism requiring its proline-rich C-terminal sequence. Moreover, we show that both proline-rich sequence and phosphorylation of Y335 of Dgk-α mediate: (i) its enzymatic activation, (ii) its ability to interact respectively with SH3 and SH2 domains of Src, (iii) its recruitment to the membrane. In addition, we show that phosphorylation of Dgk-α on Y335 is required for HGF-induced motility, while its constitutive recruitment at the membrane by myristylation is sufficient to trigger spontaneous motility in absence of HGF. Providing the first evidence that tyrosine phosphorylation of Dgk-α is required for growth-factors-induced activation and membrane recruitment, these findings underscore its relevance as a rheostat, whose activation is a threshold to elicit growth factors-induced migratory signaling.

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Acknowledgements

We thank Cecilia Deantonio, Andrea Pighini, Miriam Gaggianesi, Erika Loggia and Marianna Notario for their helpful work throughout the project. This study was supported by grants from AICR (International Agency for Cancer Research, Glasgow), the Italian Ministry for University and Research (PRIN 2004-05 and FIRB 2001 post-genomic program to AG; FIRB 2001 and RBNE019J9W_003 to OP), Regione Piemonte (Ricerca Sanitaria and CIPE), Fondazione Cariplo, AIRC (Italian Association for Cancer Research) to AG, Istituto Superiore di Sanità ‘Progetto Malattie Rare’ to OP. NF was supported by FIRB.

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Author notes

  1. G Baldanzi and S Cutrupi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Sciences, University Amedeo Avogadro of Piemonte Oriental, Novara, Italy

    G Baldanzi, F Chianale, V Gnocchi, E Rainero, P Porporato, F Sinigaglia & A Graziani

  2. Centro Di Ricerca E.Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy

    G Baldanzi & O Parolini

  3. Department of Human and Animal Biology, University of Turin, Turin, Italy

    S Cutrupi

  4. Department of Clinical and Experimental Medicine, University Amedeo Avogadro of Piemonte Orientale, Novara, Italy

    N Filigheddu

  5. Division of Cellular Biochemistry at the Netherlands Cancer Institute, Amsterdam, The Netherlands

    W J van Blitterswijk

  6. Institute for Cancer Research and Treatment (I.R.C.C.) and Department of Oncological Sciences, University of Torino, Turin, Italy

    F Bussolino

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  1. G Baldanzi
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Correspondence to A Graziani.

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Baldanzi, G., Cutrupi, S., Chianale, F. et al. Diacylglycerol kinase-α phosphorylation by Src on Y335 is required for activation, membrane recruitment and Hgf-induced cell motility. Oncogene 27, 942–956 (2008). https://doi.org/10.1038/sj.onc.1210717

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