Shear stress induced reorganization of the keratin intermediate filament network requires phosphorylation by protein kinase C zeta

Mol Biol Cell. 2009 Jun;20(11):2755-65. doi: 10.1091/mbc.e08-10-1028. Epub 2009 Apr 8.

Abstract

Keratin intermediate filaments (KIFs) form a fibrous polymer network that helps epithelial cells withstand external mechanical forces. Recently, we established a correlation between the structure of the KIF network and its local mechanical properties in alveolar epithelial cells. Shear stress applied across the cell surface resulted in the structural remodeling of KIF and a substantial increase in the elastic modulus of the network. This study examines the mechanosignaling that regulates the structural remodeling of the KIF network. We report that the shear stress-mediated remodeling of the KIF network is facilitated by a twofold increase in the dynamic exchange rate of KIF subunits, which is regulated in a PKC zeta and 14-3-3-dependent manner. PKC zeta phosphorylates K18pSer33, and this is required for the structural reorganization because the KIF network in A549 cells transfected with a dominant negative PKC zeta, or expressing the K18Ser33Ala mutation, is unchanged. Blocking the shear stress-mediated reorganization results in reduced cellular viability and increased apoptotic levels. These data suggest that shear stress mediates the phosphorylation of K18pSer33, which is required for the reorganization of the KIF network, resulting in changes in mechanical properties of the cell that help maintain the integrity of alveolar epithelial cells.

Publication types

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

MeSH terms

  • 14-3-3 Proteins / genetics
  • 14-3-3 Proteins / metabolism
  • Amino Acid Substitution
  • Cell Cycle
  • Cell Line, Tumor
  • Fluorescent Antibody Technique
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Intermediate Filaments / genetics
  • Intermediate Filaments / metabolism*
  • Keratin-18 / genetics
  • Keratin-18 / metabolism
  • Keratins / genetics
  • Keratins / metabolism*
  • Microscopy, Confocal
  • Mutation
  • Phosphorylation
  • Protein Binding
  • Protein Kinase C / metabolism*
  • Serine / genetics
  • Serine / metabolism
  • Stress, Mechanical

Substances

  • 14-3-3 Proteins
  • Keratin-18
  • Green Fluorescent Proteins
  • Serine
  • Keratins
  • protein kinase C zeta
  • Protein Kinase C