The LIM domain of zyxin is sufficient for force-induced accumulation of zyxin during cell migration

Biophys J. 2011 Sep 7;101(5):1069-75. doi: 10.1016/j.bpj.2011.08.001.


Cellular responses to mechanical perturbation are vital to cell physiology. In particular, migrating cells have been shown to sense substrate stiffness and alter cell morphology and speed. Zyxin is a focal adhesion protein that responds to external mechanical forces; however, the mechanisms of zyxin recruitment at force-bearing sites are unknown. Using force-sensing microfabricated substrates, we simultaneously measured traction force and zyxin recruitment at force-bearing sites. GFP-tagged zyxin accumulates at force-bearing sites at the leading edge, but not at the trailing edge, of migrating epithelial cells. Zyxin recruitment at force-bearing sites depends on Rho-kinase and myosin II activation, suggesting that zyxin responds not only to the externally applied force, as previously shown, but also to the internally generated actin-myosin force. Zyxin in turn recruits vasodilator-stimulated phosphoprotein, a regulator of actin assembly, to force-bearing sites. To dissect the domains of zyxin that are essential for this unique force-dependent accumulation, we generated two zyxin truncation mutants: one lacking the LIM domain (ΔLIM) and one containing only the LIM domain with all three LIM motifs (LIM). GFP-tagged ΔLIM does not localize to the force-bearing sites, but GFP-tagged zyxin LIM-domain is sufficient for the recruitment to and dynamics at force-bearing focal adhesions. Furthermore, one or two LIM motifs are not sufficient for force-dependent accumulation, suggesting that all three LIM motifs are required. Therefore, the LIM domain of zyxin recruits zyxin to force-bearing sites at the leading edge of migrating cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Animals
  • Binding Sites
  • Biomechanical Phenomena
  • Cell Adhesion Molecules / metabolism
  • Cell Line
  • Cell Movement*
  • Dogs
  • Humans
  • Mechanical Phenomena*
  • Microfilament Proteins / metabolism
  • Myosin Type II / metabolism
  • Phosphoproteins / metabolism
  • Protein Structure, Tertiary
  • Zyxin / chemistry*
  • Zyxin / metabolism*
  • rho-Associated Kinases / metabolism


  • Cell Adhesion Molecules
  • Microfilament Proteins
  • Phosphoproteins
  • Zyxin
  • vasodilator-stimulated phosphoprotein
  • rho-Associated Kinases
  • Myosin Type II