The effects of the small GTPase RhoA on the muscarinic contraction of airway smooth muscle result from its role in regulating actin polymerization

Wenwu Zhang, Liping Du, Susan J. Gunst

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

The small GTPase RhoA increases the Ca2+ sensitivity of smooth muscle contraction and myosin light chain (MLC) phosphorylation by inhibiting the activity of MLC phosphatase. RhoA is also a known regulator of cytoskeletal dynamics and actin polymerization in many cell types. In airway smooth muscle (ASM), contractile stimulation induces MLC phosphorylation and actin polymerization, which are both required for active tension generation. The objective of this study was to evaluate the primary mechanism by which RhoA regulates active tension generation in intact ASM during stimulation with acetylcholine (ACh). RhoA activity was inhibited in canine tracheal smooth muscle tissues by expressing the inactive RhoA mutant, RhoA T19N, in the intact tissues or by treating them with the cell-permeant RhoA inhibitor, exoenzyme C3 transferase. RhoA inactivation reduced ACh-induced contractile force by ∼60% and completely inhibited ACh-induced actin polymerization but inhibited ACh-induced MLC phosphorylation by only ∼20%. Inactivation of MLC phosphatase with calyculin A reversed the reduction in MLC phosphorylation caused by RhoA inactivation, but calyculin A did not reverse the depression of active tension and actin polymerization caused by RhoA inactivation. The MLC kinase inhibitor, ML-7, inhibited ACh-induced MLC phosphorylation by ∼80% and depressed active force by ∼70% but did not affect ACh-induced actin polymerization, demonstrating that ACh-stimulated actin polymerization occurs independently of MLC phosphorylation. We conclude that the RhoA-mediated regulation of ACh-induced contractile tension in ASM results from its role in mediating actin polymerization rather than from effects on MLC phosphatase or MLC phosphorylation.

Original languageEnglish (US)
Pages (from-to)C298-C306
JournalAmerican Journal of Physiology - Cell Physiology
Volume299
Issue number2
DOIs
StatePublished - Aug 2010

Keywords

  • Calcium sensitization
  • Calyculin A
  • Cytoskeleton
  • Myosin light chain kinase
  • Myosin light chain phosphatase
  • Myosin light chain phosphorylation
  • Myosin phosphatase

ASJC Scopus subject areas

  • Cell Biology
  • Physiology
  • Medicine(all)

Fingerprint Dive into the research topics of 'The effects of the small GTPase RhoA on the muscarinic contraction of airway smooth muscle result from its role in regulating actin polymerization'. Together they form a unique fingerprint.

  • Cite this