Recruitment of Dbl by ezrin and dystroglycan drives membrane proximal Cdc42 activation and filopodia formation

Cell Cycle. 2007 Feb 1;6(3):353-63. doi: 10.4161/cc.6.3.3819. Epub 2007 Feb 5.

Abstract

Dystroglycan is an essential laminin binding cell adhesion molecule, which is also an adaptor for several SH2 domain-containing signaling molecules and as a scaffold for the ERK-MAP kinase cascade. Loss of dystroglycan function is implicated in muscular dystrophies and the aetiology of epithelial cancers. We have previously demonstrated a role for dystroglycan and ezrin in the formation of filopodia structures. Here we demonstrate the existence of a dystroglycan:ezrin:Dbl complex that is targeted to the membrane by dystroglycan where it drives local Cdc42 activation and the formation of filopodia. Deletion of an ezrin binding site in dystroglycan prevented the association with ezrin and Dbl and the formation of filopodia. Furthermore, expression of the dystroglycan cytoplasmic domain alone had a dominant-negative effect on filopodia formation and Cdc42 activation by sequestering ezrin and Dbl away from the membrane. Depletion of dystroglycan inhibited Cdc42-induced filopodia formation. For the first time we also demonstrate co-localization of Cdc42 and dystroglycan at the tips of dynamic filopodia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Binding Sites
  • COS Cells
  • Cell Line
  • Chlorocebus aethiops
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism*
  • Dystroglycans / genetics
  • Dystroglycans / metabolism*
  • Fluorescent Antibody Technique
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Immunoblotting
  • Immunoprecipitation
  • Mice
  • Mutation
  • Protein Binding
  • Pseudopodia / metabolism*
  • RNA Interference
  • Rats
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Swiss 3T3 Cells
  • cdc42 GTP-Binding Protein / genetics
  • cdc42 GTP-Binding Protein / metabolism*
  • rho GTP-Binding Proteins / metabolism

Substances

  • Actins
  • Cytoskeletal Proteins
  • Guanine Nucleotide Exchange Factors
  • Recombinant Fusion Proteins
  • ezrin
  • Dystroglycans
  • Green Fluorescent Proteins
  • cdc42 GTP-Binding Protein
  • rho GTP-Binding Proteins