Pyk2 regulates cell-edge protrusion dynamics by interacting with Crk

Mol Biol Cell. 2021 Nov 1;32(21):ar17. doi: 10.1091/mbc.E20-10-0640. Epub 2021 Aug 25.

Abstract

Focal adhesion kinase (FAK) is well established as a regulator of cell migration, but whether and how the closely related proline-rich tyrosine kinase 2 (Pyk2) regulates fibroblast motility is still under debate. Using mouse embryonic fibroblasts (MEFs) from Pyk2-/- mice, we show here, for the first time, that lack of Pyk2 significantly impairs both random and directed fibroblast motility. Pyk2-/- MEFs show reduced cell-edge protrusion dynamics, which is dependent on both the kinase and protein-protein binding activities of Pyk2. Using bioinformatics analysis of in vitro high- throughput screens followed by text mining, we identified CrkI/II as novel substrates and interactors of Pyk2. Knockdown of CrkI/II shows altered dynamics of cell-edge protrusions, which is similar to the phenotype observed in Pyk2-/- MEFs. Moreover, epistasis experiments suggest that Pyk2 regulates the dynamics of cell-edge protrusions via direct and indirect interactions with Crk that enable both activation and down-regulation of Crk-mediated cytoskeletal signaling. This complex mechanism may enable fine-tuning of cell-edge protrusion dynamics and consequent cell migration on the one hand together with tight regulation of cell motility, a process that should be strictly limited to specific time and context in normal cells, on the other hand.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Movement / genetics*
  • Cell Movement / physiology
  • Cell Surface Extensions / metabolism
  • Cytoskeleton / metabolism
  • Fibroblasts / metabolism*
  • Focal Adhesion Kinase 2 / metabolism*
  • Focal Adhesion Kinase 2 / physiology
  • Mice
  • Mitogen-Activated Protein Kinases / metabolism
  • Phosphorylation
  • Protein Binding
  • Proto-Oncogene Proteins c-crk / genetics
  • Proto-Oncogene Proteins c-crk / metabolism
  • Signal Transduction

Substances

  • Proto-Oncogene Proteins c-crk
  • Focal Adhesion Kinase 2
  • Ptk2b protein, mouse
  • Mitogen-Activated Protein Kinases