The small GTPase Cdc42 regulates actin polymerization and tension development during contractile stimulation of smooth muscle

J Biol Chem. 2004 Dec 10;279(50):51722-8. doi: 10.1074/jbc.M408351200. Epub 2004 Sep 27.


Contractile stimulation induces actin polymerization in smooth muscle tissues and cells, and the inhibition of actin polymerization depresses smooth muscle force development. In the present study, the role of Cdc42 in the regulation of actin polymerization and tension development in smooth muscle was evaluated. Acetylcholine stimulation of tracheal smooth muscle tissues increased the activation of Cdc42. Plasmids encoding wild type Cdc42 or a dominant negative Cdc42 mutant, Asn-17 Cdc42, were introduced into tracheal smooth muscle strips by reversible permeabilization, and tissues were incubated for 2 days to allow for protein expression. Expression of recombinant proteins was confirmed by immunoblot analysis. The expression of the dominant negative Cdc42 mutant inhibited contractile force and the increase in actin polymerization in response to acetylcholine stimulation but did not inhibit the increase in myosin light chain phosphorylation. The expression of wild type Cdc42 had no significant effect on force, actin polymerization, or myosin light chain phosphorylation. Contractile stimulation increased the association of neuronal Wiskott-Aldrich syndrome protein with Cdc42 and the Arp2/3 (actin-related protein) complex in smooth muscle tissues expressing wild type Cdc42. The agonist-induced increase in these protein interactions was inhibited in tissues expressing the inactive Cdc42 mutant. We conclude that Cdc42 activation regulates active tension development and actin polymerization during contractile stimulation. Cdc42 may regulate the activation of neuronal Wiskott-Aldrich syndrome protein and the actin related protein complex, which in turn regulate actin filament polymerization initiated by the contractile stimulation of smooth muscle.

Publication types

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

MeSH terms

  • Acetylcholine / pharmacology
  • Actin-Related Protein 2
  • Actins / chemistry
  • Actins / metabolism*
  • Animals
  • Cytoskeletal Proteins / metabolism
  • Dogs
  • Enzyme Activation / drug effects
  • Humans
  • In Vitro Techniques
  • Muscle Contraction / physiology*
  • Muscle, Smooth / drug effects
  • Muscle, Smooth / physiology*
  • Mutagenesis, Site-Directed
  • Myosin Light Chains / metabolism
  • Nerve Tissue Proteins / metabolism
  • Phosphorylation
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Trachea / physiology
  • Wiskott-Aldrich Syndrome Protein, Neuronal
  • cdc42 GTP-Binding Protein / genetics
  • cdc42 GTP-Binding Protein / metabolism*


  • ACTR2 protein, human
  • Actin-Related Protein 2
  • Actins
  • Cytoskeletal Proteins
  • Myosin Light Chains
  • Nerve Tissue Proteins
  • Recombinant Proteins
  • WASL protein, human
  • Wiskott-Aldrich Syndrome Protein, Neuronal
  • cdc42 GTP-Binding Protein
  • Acetylcholine