Mechanosensing Machinery for Cells Under Low Substratum Rigidity

Am J Physiol Cell Physiol. 2008 Dec;295(6):C1579-89. doi: 10.1152/ajpcell.00223.2008. Epub 2008 Oct 15.

Abstract

Mechanical stimuli are essential during development and tumorigenesis. However, how cells sense their physical environment under low rigidity is still unknown. Here we show that low rigidity of collagen gel downregulates beta(1)-integrin activation, clustering, and focal adhesion kinase (FAK) Y397 phosphorylation, which is mediated by delayed raft formation. Moreover, overexpression of autoclustered beta(1)-integrin (V737N), but not constitutively active beta(1)-integrin (G429N), rescues FAKY397 phosphorylation level suppressed by low substratum rigidity. Using fluorescence resonance energy transfer to assess beta(1)-integrin clustering, we have found that substratum rigidity between 58 and 386 Pa triggers beta(1)-integrin clustering in a dose-dependent manner, which is highly dependent on actin filaments but not microtubules. Furthermore, augmentation of beta(1)-integrin clustering enhances the interaction between beta(1)-integrin, FAK, and talin. Our results indicate that contact with collagen fibrils is not sufficient for integrin activation. However, substratum rigidity is required for integrin clustering and activation. Together, our findings provide new insight into the mechanosensing machinery and the mode of action for epithelial cells in response to their physical environment under low rigidity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Animals
  • Blotting, Western
  • Cell Line
  • Collagen
  • Dogs
  • Epithelial Cells / physiology*
  • Extracellular Matrix / metabolism*
  • Fluorescence Resonance Energy Transfer
  • Focal Adhesion Protein-Tyrosine Kinases
  • Integrin beta1 / metabolism*
  • Mechanotransduction, Cellular / physiology*
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Phosphorylation
  • Protein Transport / physiology
  • Signal Transduction / physiology*

Substances

  • Integrin beta1
  • Collagen
  • Focal Adhesion Protein-Tyrosine Kinases