H-Ras activation promotes cytoplasmic accumulation and phosphoinositide 3-OH kinase association of beta-catenin in epidermal keratinocytes

J Cell Biol. 1999 Sep 6;146(5):967-80. doi: 10.1083/jcb.146.5.967.

Abstract

The mechanisms underlying downregulation of the cadherin/catenin complexes and beta-catenin signaling during tumor progression are not fully understood. We have analyzed the effect of oncogenic H-Ras on E-cadherin/catenin complex formation/stabilization and beta-catenin distribution in epidermal keratinocytes. Microinjection or stable expression of V12Ras into keratinocytes promotes the loss of E-cadherin and alpha-catenin and relocalization of beta-catenin to the cytoplasm and nucleus. Moreover, these effects are dependent on PI3K (phosphoinositide 3-OH kinase) activity. Interestingly, a strong association of p85alpha and p110alpha subunits of PI3K with beta-catenin is induced in V12Ras-expressing keratinocytes, and in vitro binding assays show a direct interaction between beta-catenin and p85alpha. Overexpression of either V12Ras or constitutively active p110alpha induces metabolic stabilization of beta-catenin and promotes its accumulation in cytoplasmic and nuclear pools. In addition, the interaction of beta-catenin with the adenomatous polyposis coli protein is blocked in V12Ras and p110alpha transformants though no changes in glycogen synthase kinase 3 beta activity could be detected. Nevertheless, in V12Ras transformants the in vivo phosphorylation of beta-catenin in Ser residues is strongly decreased. These results indicate that H-Ras activation induces the relocalization and cytoplasmic stabilization of beta-catenin by a mechanism involving its interaction with PI3K.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenomatous Polyposis Coli Protein
  • Animals
  • Cadherins / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Cell Line
  • Cell Membrane / metabolism
  • Cell Nucleus / metabolism
  • Cell Transformation, Neoplastic
  • Cytoplasm / metabolism*
  • Cytoskeletal Proteins / metabolism*
  • Enzyme Activation
  • Glycogen Synthase Kinase 3
  • Glycogen Synthase Kinases
  • Keratinocytes / cytology
  • Keratinocytes / enzymology
  • Keratinocytes / metabolism*
  • Mice
  • Microinjections
  • Oncogene Protein p21(ras) / genetics
  • Oncogene Protein p21(ras) / metabolism*
  • Phosphatidylinositol 3-Kinases / chemistry
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphoinositide-3 Kinase Inhibitors
  • Phosphorylation
  • Phosphoserine / analysis
  • Phosphoserine / metabolism
  • Phosphotyrosine / analysis
  • Protein Binding
  • Trans-Activators*
  • alpha Catenin
  • beta Catenin

Substances

  • Adenomatous Polyposis Coli Protein
  • CTNNB1 protein, mouse
  • Cadherins
  • Ctnna1 protein, mouse
  • Cytoskeletal Proteins
  • Phosphoinositide-3 Kinase Inhibitors
  • Trans-Activators
  • alpha Catenin
  • beta Catenin
  • Phosphoserine
  • Phosphotyrosine
  • Glycogen Synthase Kinases
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Glycogen Synthase Kinase 3
  • Oncogene Protein p21(ras)