Modulation of FAK and Src adhesion signaling occurs independently of adhesion complex composition

J Cell Biol. 2016 Feb 1;212(3):349-64. doi: 10.1083/jcb.201508080.

Abstract

Integrin adhesion complexes (IACs) form mechanochemical connections between the extracellular matrix and actin cytoskeleton and mediate phenotypic responses via posttranslational modifications. Here, we investigate the modularity and robustness of the IAC network to pharmacological perturbation of the key IAC signaling components focal adhesion kinase (FAK) and Src. FAK inhibition using AZ13256675 blocked FAK(Y397) phosphorylation but did not alter IAC composition, as reported by mass spectrometry. IAC composition was also insensitive to Src inhibition using AZD0530 alone or in combination with FAK inhibition. In contrast, kinase inhibition substantially reduced phosphorylation within IACs, cell migration and proliferation. Furthermore using fluorescence recovery after photobleaching, we found that FAK inhibition increased the exchange rate of a phosphotyrosine (pY) reporter (dSH2) at IACs. These data demonstrate that kinase-dependent signal propagation through IACs is independent of gross changes in IAC composition. Together, these findings demonstrate a general separation between the composition of IACs and their ability to relay pY-dependent signals.

Publication types

  • Research Support, Non-U.S. Gov't
  • Webcast

MeSH terms

  • Animals
  • Cell Adhesion / drug effects*
  • Cell Movement
  • Cell Proliferation
  • Dose-Response Relationship, Drug
  • Fibroblasts / drug effects
  • Fibroblasts / enzymology*
  • Focal Adhesion Kinase 1 / antagonists & inhibitors
  • Focal Adhesion Kinase 1 / genetics
  • Focal Adhesion Kinase 1 / metabolism*
  • Focal Adhesions / drug effects
  • Focal Adhesions / enzymology*
  • Focal Adhesions / genetics
  • HEK293 Cells
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Phosphorylation
  • Phosphotyrosine / metabolism
  • Protein Binding
  • Protein Kinase Inhibitors / pharmacology
  • Signal Transduction / drug effects*
  • Time Factors
  • Transfection
  • src Homology Domains
  • src-Family Kinases / antagonists & inhibitors
  • src-Family Kinases / metabolism*

Substances

  • Protein Kinase Inhibitors
  • Phosphotyrosine
  • Focal Adhesion Kinase 1
  • PTK2 protein, human
  • src-Family Kinases