Expression of BRG1, a human SWI/SNF component, affects the organisation of actin filaments through the RhoA signalling pathway

J Cell Sci. 2002 Jul 1;115(Pt 13):2735-46. doi: 10.1242/jcs.115.13.2735.

Abstract

The human BRG1 (brahma-related gene 1) protein is a component of the SWI/SNF family of the ATP-dependent chromatin remodelling complexes. We show here that expression of the BRG1 protein, but not of an ATPase-deficient BRG1 protein, in BRG1-deficient SW13 cells alters the organisation of actin filaments. BRG1 expression induces the formation of thick actin filament bundles resembling stress-fibres, structures that are rarely seen in native SW13 cells. BRG1 expression does not influence the activity state of the RhoA-GTPase, which is involved in stress-fibre formation. We find that RhoA is equally activated by stimuli, such as serum, in BRG1-expressing cells, ATPase-deficient BRG1-expressing cells and native SW13 cells. However, the activation of RhoA by lysophosphatidic acid and serum does not trigger the formation of stress-fibre-like structures in SW13 cells. Activation of the RhoA-GTPase in BRG1-expressing cells induces stress-fibre-like structures, indicating that the BRG1 can couple RhoA activation to stress-fibre formation. At least two downstream effectors are involved in stress-fibre formation, Rho-kinase/ROCK and Dia. BRG1 expression, but not the expression of the ATP-deficient BRG1, increases the protein level of ROCK1, one form of the Rho-kinase/ROCK. That this is of importance is supported by the findings that an increased Rho-kinase/ROCK activity in SW13 cells, obtained by overexpressing wild-type ROCK1 and ROCK2, induces stress-fibre formation. No specificity between the two Rho-kinase/ROCK forms exists. Our results suggest that the BRG1 protein affects the RhoA pathway by increasing the protein level of ROCK1, which allows stress-fibre-like structures to form.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism*
  • Animals
  • Cell Size / genetics
  • Cells, Cultured
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA Helicases
  • Eukaryotic Cells / cytology
  • Eukaryotic Cells / metabolism*
  • Fluorescent Antibody Technique
  • Gene Expression Regulation / physiology
  • Genes, Regulator / genetics
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Reaction Time / genetics
  • Signal Transduction / genetics
  • Stress, Mechanical
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Up-Regulation / genetics
  • rho-Associated Kinases
  • rhoA GTP-Binding Protein / genetics
  • rhoA GTP-Binding Protein / metabolism*

Substances

  • Chromosomal Proteins, Non-Histone
  • Intracellular Signaling Peptides and Proteins
  • Nuclear Proteins
  • SWI-SNF-B chromatin-remodeling complex
  • Transcription Factors
  • Protein Serine-Threonine Kinases
  • ROCK1 protein, human
  • ROCK2 protein, human
  • rho-Associated Kinases
  • SMARCA4 protein, human
  • DNA Helicases
  • rhoA GTP-Binding Protein