Phosphorylation of Tyrosine Residues 31 and 118 on Paxillin Regulates Cell Migration through an Association with CRK in NBT-II Cells

• Published in: The Journal of cell biology 2000-03, Vol.148 (5), p.957-969
• Main Authors: Petit, Valérie, Boyer, Brigitte, Lentz, Delphine, Turner, Christopher E., Thiery, Jean Paul, Vallés, Ana M.
• Format: Article
• Language: English
• Subjects: Amino Acid Substitution; Animals; Antibodies; Binding Sites - genetics; Cell Adhesion; Cell Adhesion Molecules - metabolism; Cell lines; Cell motility; Cell Movement - genetics; Cells; Clone Cells; Collagen - metabolism; Collagen - pharmacology; Collagens; Cytoskeletal Proteins - genetics; Cytoskeletal Proteins - metabolism; Epithelial cells; Focal Adhesion Kinase 1; Focal Adhesion Protein-Tyrosine Kinases; Focal adhesions; Molecular biology; Mutagenesis, Site-Directed; Mutation; Original; Paxillin; Phosphoproteins - genetics; Phosphoproteins - metabolism; Phosphorylation; Phosphorylation - drug effects; Protein Binding - genetics; Protein Kinases - genetics; Protein Kinases - metabolism; Protein-Tyrosine Kinases - metabolism; Proteins; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-crk; Rats; src Homology Domains - genetics; Swiss 3T3 cells; Tumor Cells, Cultured; Tyrosine - metabolism; Urinary Bladder Neoplasms - metabolism; Urinary Bladder Neoplasms - pathology
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.148.5.957

Identification of signaling molecules that regulate cell migration is important for understanding fundamental processes in development and the origin of various pathological conditions. The migration of Nara Bladder Tumor II (NBT-II) cells was used to determine which signaling molecules are specifically involved in the collagen-mediated locomotion. We show here that paxillin is tyrosine phosphorylated after induction of motility on collagen. Overexpression of paxillin mutants in which tyrosine 31 and/or tyrosine 118 were replaced by phenylalanine effectively impaired cell motility. Moreover, stimulation of motility by collagen preferentially enhanced the association of paxillin with the SH2 domain of the adaptor protein CrkII. Mutations in both tyrosine 31 and 118 diminished the phosphotyrosine content of paxillin and prevented the formation of the paxillin-Crk complex, suggesting that this association is necessary for collagen-mediated NBT-II cell migration. Other responses to collagen, such as cell adhesion and spreading, were not affected by these mutations. Overexpression of wild-type paxillin or Crk could bypass the migration-deficient phenotype. Both the SH2 and the SH3 domains of CrkII are shown to play a critical role in this collagen-mediated migration. These results demonstrate the important role of the paxillin-Crk complex in the collagen-induced cell motility.