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.
%0 Journal Article
%1 Eiseler2010
%A Eiseler, Tim
%A Hausser, Angelika
%A De Kimpe, Line
%A Van Lint, Johan
%A Pfizenmaier, Klaus
%D 2010
%J Journal of Biological Chemistry
%K 2010 hausser izi pfizenmaier
%N 24
%P 18672--18683
%R 10.1074/jbc.M109.093880
%T Protein kinase D controls actin polymerization and cell motility through phosphorylation of cortactin
%U https://www.ncbi.nlm.nih.gov/pubmed/20363754
%V 285
%X 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.
%Z Eiseler, TimHausser, AngelikaDe Kimpe, LineVan Lint, JohanPfizenmaier, KlausengResearch Support, Non-U.S. Gov'tJ Biol Chem. 2010 Jun 11;285(24):18672-83. doi: 10.1074/jbc.M109.093880. Epub 2010 Apr 2.
%7 2010/04/07
%@ 1083-351X (Electronic)$\backslash$r0021-9258 (Linking)
@article{Eiseler2010,
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.},
added-at = {2018-02-01T15:26:42.000+0100},
annote = {Eiseler, TimHausser, AngelikaDe Kimpe, LineVan Lint, JohanPfizenmaier, KlausengResearch Support, Non-U.S. Gov'tJ Biol Chem. 2010 Jun 11;285(24):18672-83. doi: 10.1074/jbc.M109.093880. Epub 2010 Apr 2.},
author = {Eiseler, Tim and Hausser, Angelika and {De Kimpe}, Line and {Van Lint}, Johan and Pfizenmaier, Klaus},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2a5b17d8c33dfcd7c0aa816f443836692/cristiano},
doi = {10.1074/jbc.M109.093880},
edition = {2010/04/07},
interhash = {74449fff417cce16821e78ce54247b7e},
intrahash = {a5b17d8c33dfcd7c0aa816f443836692},
isbn = {1083-351X (Electronic)$\backslash$r0021-9258 (Linking)},
issn = {00219258},
journal = {Journal of Biological Chemistry},
keywords = {2010 hausser izi pfizenmaier},
number = 24,
pages = {18672--18683},
pmid = {20363754},
timestamp = {2018-07-25T12:24:13.000+0200},
title = {{Protein kinase D controls actin polymerization and cell motility through phosphorylation of cortactin}},
url = {https://www.ncbi.nlm.nih.gov/pubmed/20363754},
volume = 285,
year = 2010
}