Temporal dissection of beta1-integrin signaling indicates a role for p130Cas-Crk in filopodia formation

J Biol Chem. 2004 May 28;279(22):22893-901. doi: 10.1074/jbc.M309693200. Epub 2004 Mar 24.

Abstract

Invasin-promoted spreading of beta1-integrin-deficient cells, transfected with the beta1A- or beta1B-integrin splice variants, were used to dissect early beta1-integrin signaling events. The beta1B isoform, which has a different membrane-distal part of the cytoplasmic tail from beta1A, is defective in signaling and function. When plated on surfaces coated with the high affinity ligand invasin, beta1B-integrin-expressing cells spread by forming filopodia with distinct adhesive phosphotyrosine complexes at the tips, without signs of lamellipodia. This suggested that the beta1B-integrin mediated a partial signaling sufficient for formation of filopodia but insufficient for lamellipodia formation. When screening for proteins present in the distal filopodial phosphotyrosine complexes of beta1B cells, p130Cas and the filopodia proteins vasodilator-stimulated phosphoprotein and talin were found, whereas the typical focal complex proteins focal adhesion kinase, paxillin, and vinculin were not. Invasin-promoted adhesion induced complex formation of p130Cas and the adapter Crk. Moreover, Crk together with Dock180 were present at the filopodial tips of beta1B-integrin-expressing cells, and there was a prominent Rac1 activation. Expression of dominant negative variants of p130Cas or CrkII blocked beta1B-integrin-mediated filopodia formation, indicating that this signaling scaffold is central in this process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Crk-Associated Substrate Protein
  • Integrin beta1 / physiology*
  • Phosphorylation
  • Proteins / physiology*
  • Proto-Oncogene Proteins / physiology
  • Proto-Oncogene Proteins c-crk
  • Pseudopodia / physiology*
  • Rabbits
  • Rats
  • Retinoblastoma-Like Protein p130
  • Signal Transduction

Substances

  • Bcar1 protein, rat
  • Crk protein, rat
  • Crk-Associated Substrate Protein
  • Integrin beta1
  • Proteins
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-crk
  • Retinoblastoma-Like Protein p130