GEF-H1 is involved in agonist-induced human pulmonary endothelial barrier dysfunction

Anna A. Birukova, Djanybek Adyshev, Boris A Gorshkov, Gary M. Bokoch, Konstantin G. Birukov, Alexander Dmitriyevich Verin

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

Endothelial cell (EC) permeability is precisely controlled by cytoskeletal elements [actin filaments, microtubules (MT), intermediate filaments] and cell contact protein complexes (focal adhesions, adherens junctions, tight junctions). We have recently shown that the edemagenic agonist thrombin caused partial MT disassembly, which was linked to activation of small GTPase Rho, Rho-mediated actin remodeling, cell contraction, and dysfunction of lung EC barrier. GEF-H1 is an MT-associated Rho-specific guanosine nucleotide (GDP/GTP) exchange factor, which in MT-unbound state stimulates Rho activity. In this study we tested hypothesis that GEF-H1 may be a key molecule involved in Rho activation, myosin light chain phosphorylation, actin remodeling, and EC barrier dysfunction associated with partial MT disassembly. Our results show that depletion of GEF-H1 or expression of dominant negative GEF-H1 mutant significantly attenuated permeability increase, actin stress fiber formation, and increased MLC and MYPT1 phosphorylation induced by thrombin or MT-depolymerizing agent nocodazole. In contrast, expression of wild-type or activated GEF-H1 mutants dramatically enhanced thrombin and nocodazole effects on stress fiber formation and cell retraction. These results show a critical role for the GEF-H1 in the Rho activation caused by MT disassembly and suggest GEF-H1 as a key molecule involved in cross talk between MT and actin cytoskeleton in agonist-induced Rho-dependent EC barrier regulation.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume290
Issue number3
DOIs
StatePublished - Mar 1 2006

Fingerprint

Microtubules
Lung
Endothelial Cells
Thrombin
Nocodazole
Actins
Stress Fibers
Actin Cytoskeleton
Permeability
Phosphorylation
Guanine Nucleotide Exchange Factors
Adherens Junctions
Myosin Light Chains
Focal Adhesions
Monomeric GTP-Binding Proteins
Intermediate Filaments
Guanosine
Tight Junctions
Guanosine Triphosphate
Nucleotides

Keywords

  • Cytoskeleton
  • Guanine nucleotide exchange factor
  • Microtubules
  • Permeability
  • Pulmonary endothelium
  • Thrombin

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

Cite this

GEF-H1 is involved in agonist-induced human pulmonary endothelial barrier dysfunction. / Birukova, Anna A.; Adyshev, Djanybek; Gorshkov, Boris A; Bokoch, Gary M.; Birukov, Konstantin G.; Verin, Alexander Dmitriyevich.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 290, No. 3, 01.03.2006.

Research output: Contribution to journalArticle

Birukova, Anna A. ; Adyshev, Djanybek ; Gorshkov, Boris A ; Bokoch, Gary M. ; Birukov, Konstantin G. ; Verin, Alexander Dmitriyevich. / GEF-H1 is involved in agonist-induced human pulmonary endothelial barrier dysfunction. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2006 ; Vol. 290, No. 3.
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