title: Cyclic GMP-dependent protein kinase signaling pathway inhibits RhoA-induced Ca2+ sensitization of contraction in vascular smooth muscle.
abstract: The potent vasodilator action of cyclic GMP-dependent protein kinase (cGK) involves decreasing the Ca(2+) sensitivity of contraction of smooth muscle via stimulation of myosin light chain phosphatase through unknown mechanisms (Wu, X., Somlyo, A. V., and Somlyo, A. P. (1996) Biochem. Biophys. Res. Commun. 220, 658-663). Myosin light chain phosphatase activity is controlled by the small GTPase RhoA and its target Rho kinase. Here we demonstrate cGMP effects mediated by cGK that inhibit RhoA-dependent Ca(2+) sensitization of contraction of blood vessels and actin cytoskeleton organization in cultured vascular myocytes. Ca(2+) sensitization and actin organization were inhibited by both 8-bromo-cGMP and sodium nitroprusside (SNP). SNP also caused translocation of activated RhoA from the membrane to the cytosol. SNP-induced actin disassembly was lost in vascular myocytes in culture after successive passages but was restored by transfection of cells with cGK I. Furthermore, cGK phosphorylated RhoA in vitro, and addition of cGK I inhibited RhoA-induced Ca(2+) sensitization in permeabilized smooth muscle. 8-Bromo-cGMP-induced actin disassembly was inhibited in vascular myocytes expressing RhoA(Ala-188), a mutant that could not be phosphorylated. Collectively, these results indicate that cGK phosphorylates and inhibits RhoA and suggest that the consequent inhibition of RhoA-induced Ca(2+) sensitization and actin cytoskeleton organization contributes to the vasodilator action of nitric oxide.
authors.affiliation: Laboratoire de Physiologie Cellulaire et Moléculaire, INSERM U-533, Faculté des Sciences, 44322 Nantes, France.
authors.forename: V
authors.initials: V
authors.lastname: Sauzeau
authors.forename: H
authors.initials: H
authors.lastname: Le Jeune
authors.forename: C
authors.initials: C
authors.lastname: Cario-Toumaniantz
authors.forename: A
authors.initials: A
authors.lastname: Smolenski
authors.forename: S M
authors.initials: SM
authors.lastname: Lohmann
authors.forename: J
authors.initials: J
authors.lastname: Bertoglio
authors.forename: P
authors.initials: P
authors.lastname: Chardin
authors.forename: P
authors.initials: P
authors.lastname: Pacaud
authors.forename: G
authors.initials: G
authors.lastname: Loirand
mesh_terms: D000199:Actins / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000502:physiology; D002118:Calcium / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology; D005711:Gallopamil / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007537:Isometric Contraction / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000502:physiology*; D009599:Nitroprusside / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology; D010766:Phosphorylation; D011169:Portal Vein / Q000502:physiology; D011651:Pulmonary Artery / Q000502:physiology; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D019284:Thapsigargin / Q000494:pharmacology; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000199:Actins; D009599:Nitroprusside; D010656:Phenylephrine; C016276:8-bromocyclic GMP; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D005711:Gallopamil; D019284:Thapsigargin; D017869:Cyclic GMP-Dependent Protein Kinases; D020742:rhoA GTP-Binding Protein; D006152:Cyclic GMP; D002118:Calcium