Protein Kinase D Controls Actin Polymerization and Cell Motility through Phosphorylation of Cortactin |
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Authors: | Tim Eiseler Angelika Hausser Line De Kimpe Johan Van Lint Klaus Pfizenmaier |
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Affiliation: | From the ‡Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart 70569, Germany and ;the §Department of Molecular Cell Biology, Faculty of Medicine, K.U. Leuven, Campus Gasthuisberg O&N 1, Herestraat 49 bus 901, 3000 Leuven, Belgium |
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Abstract: | We here identify protein kinase D (PKD) as an upstream regulator of the F-actin-binding protein cortactin and the Arp actin polymerization machinery. PKD phosphorylates cortactin in vitro and in vivo at serine 298 thereby generating a 14-3-3 binding motif. In vitro, a phosphorylation-deficient cortactin-S298A protein accelerated VCA-Arp-cortactin-mediated synergistic actin polymerization and showed reduced F-actin binding, indicative of enhanced turnover of nucleation complexes. In vivo, cortactin co-localized with the nucleation promoting factor WAVE2, essential for lamellipodia extension, in the actin polymerization zone in Heregulin-treated MCF-7 cells. Using a 3-dye FRET-based approach we further demonstrate that WAVE2-Arp and cortactin prominently interact at these structures. Accordingly, cortactin-S298A significantly enhanced lamellipodia extension and directed cell migration. Our data thus unravel a previously unrecognized mechanism by which PKD controls cancer cell motility. |
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Keywords: | Actin Cell Migration Cell Motility Protein Kinases Protein Phosphorylation Arp WAVE Cortactin |
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