Chemical anoxia of tubular cells induces activation of c-Src and its translocation to the zonula adherens

Am J Physiol Renal Physiol. 2003 Mar;284(3):F488-97. doi: 10.1152/ajprenal.00172.2002. Epub 2002 Nov 5.

Abstract

Cyanide (CN)-induced chemical anoxia of cultured mouse proximal tubular (MPT) cells increased the kinase activity of c-Src by approximately threefold. 4-Amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2), a specific inhibitor of c-Src, prevented Src activation. CN also increased the permeability of MPT cell monolayers, an event ameliorated by PP2. During CN treatment, the proteins of the zonula adherens (ZA; E-cadherin and the catenins) disappeared from their normal location at cell-cell borders and appeared within the cytosol. CN also resulted in the appearance of c-Src at cell-cell borders. PP2 prevented these CN-induced alterations in the distribution of ZA proteins and c-Src. CN also increased the association of c-Src with beta-catenin and p120 and induced a substantial increase in tyrosine phosphorylation of both catenins. PP2 prevented the CN-induced phosphorylation of these catenins. In summary, we show that CN-induced chemical anoxia activates c-Src and induces its translocation to cell-cell junctions where it binds to and phosphorylates beta-catenin and p120. Our findings suggest that these events contribute to the loss of the epithelial barrier function associated with chemical anoxia.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actins / metabolism
  • Adenosine Triphosphate / metabolism
  • Adherens Junctions / metabolism*
  • Animals
  • CSK Tyrosine-Protein Kinase
  • Cadherins / metabolism
  • Catenins
  • Cell Adhesion Molecules / metabolism
  • Cell Hypoxia / drug effects
  • Cell Hypoxia / physiology
  • Cell Line
  • Cytoskeletal Proteins / metabolism
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Focal Adhesion Kinase 1
  • Focal Adhesion Protein-Tyrosine Kinases
  • Kidney Tubules, Proximal / cytology
  • Kidney Tubules, Proximal / drug effects
  • Kidney Tubules, Proximal / metabolism*
  • Mice
  • Mice, Transgenic
  • Permeability / drug effects
  • Phosphoproteins / metabolism
  • Phosphorylation / drug effects
  • Protein Transport / drug effects
  • Protein-Tyrosine Kinases*
  • Proto-Oncogene Proteins pp60(c-src) / metabolism*
  • Pyrimidines / pharmacology
  • Sodium Cyanide / pharmacology
  • Trans-Activators / metabolism
  • beta Catenin
  • src-Family Kinases

Substances

  • AG 1879
  • Actins
  • CTNNB1 protein, mouse
  • Cadherins
  • Catenins
  • Cell Adhesion Molecules
  • Cytoskeletal Proteins
  • Enzyme Inhibitors
  • Phosphoproteins
  • Pyrimidines
  • Trans-Activators
  • beta Catenin
  • delta catenin
  • Adenosine Triphosphate
  • Protein-Tyrosine Kinases
  • CSK Tyrosine-Protein Kinase
  • Focal Adhesion Kinase 1
  • Focal Adhesion Protein-Tyrosine Kinases
  • Proto-Oncogene Proteins pp60(c-src)
  • Ptk2 protein, mouse
  • src-Family Kinases
  • Sodium Cyanide