title: Ras, Rac1, and phosphatidylinositol-3-kinase (PI3K) signaling in nitric oxide induced endothelial cell migration.
abstract: The small GTP-binding proteins Ras and Rac1 are molecular switches exchanging GDP for GTP and converting external signals in response to a variety of stimuli. Ras and Rac1 play an important role in cell proliferation, cell differentiation, and cell migration. Rac1 is directly involved in the reorganization and changes in the cytoskeleton during cell motility. Nitric oxide (NO) stimulates the Ras - ERK1/2 MAP kinases signaling pathway and is involved in the interaction between Ras and the phosphatidyl-inositol-3 Kinase (PI3K) signaling pathway and cell migration. This study utilizes bradykinin (BK), which promotes endogenous production of NO, in an investigation of the role of NO in the activation of Rac1 in rabbit aortic endothelial cells (RAEC). NO-derived from BK stimulation of RAEC and incubation of the cells with the s-nitrosothiol S-nitrosoglutathione (GSNO) activated Rac1. NO-derived from BK stimulation promoted RAEC migration over a period of 12 h. The use of RAEC permanently transfected with the dominant negative mutant of Ras (Ras(N17)) or with the non-nitrosatable mutant of Ras (Ras(C118S)); and the use of specific inhibitors of: Ras, PI3K, and Rac1 resulted in inhibition of NO-mediated Rac1 activation. BK-stimulated s-nitrosylation of Ras in RAEC mediates Rac1 activation and cell migration. Inhibition of NO-mediated Rac1 activation resulted in inhibition of endothelial cell migration. In conclusion, the NO indirect activation of Rac1 involves the direct participation of Ras and PI3K in the migration of endothelial cells stimulated with BK.
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Roberta
authors.initials: R
authors.lastname: Eller-Borges
authors.affiliation: Department of Biological Sciences, Universidade Federal de São Paulo/Campus Diadema, SP, Brazil.
authors.forename: Wagner L
authors.initials: WL
authors.lastname: Batista
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Paulo E
authors.initials: PE
authors.lastname: da Costa
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Rita
authors.initials: R
authors.lastname: Tokikawa
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Marli F
authors.initials: MF
authors.lastname: Curcio
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Scheilla T
authors.initials: ST
authors.lastname: Strumillo
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Adriano
authors.initials: A
authors.lastname: Sartori
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Miriam S
authors.initials: MS
authors.lastname: Moraes
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Graciele A
authors.initials: GA
authors.lastname: de Oliveira
authors.affiliation: Department of Surgery, Escola Paulista de Medicina/Universidade Federal de São Paulo, SP, Brazil.
authors.forename: Murched O
authors.initials: MO
authors.lastname: Taha
authors.affiliation: Department of Medicine, Institute for Transformative Molecular Medicine, Case Western University, Cleveland, OH, USA.
authors.forename: Fábio V
authors.initials: FV
authors.lastname: Fonseca
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY, USA; Escuela de Medicina, Universidad Espíritu Santo, Guayaquil, Ecuador. Electronic address: Arnold.Stern@nyumc.org.
authors.forename: Arnold
authors.initials: A
authors.lastname: Stern
authors.affiliation: Department of Biochemistry, Center for Cellular and Molecular Therapy-CTCMOL, Escola Paulista de Medicina /Universidade Federal de São Paulo, SP, Brazil. Electronic address: hugo.monteiro@pesquisador.cnpq.br.
authors.forename: Hugo P
authors.initials: HP
authors.lastname: Monteiro
mesh_terms: D001920:Bradykinin / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects*; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D009569:Nitric Oxide / Q000096:biosynthesis / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D015398:Signal Transduction; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*; D018631:ras Proteins / Q000378:metabolism*
chemical_list: D009569:Nitric Oxide; D019869:Phosphatidylinositol 3-Kinases; D020830:rac1 GTP-Binding Protein; D018631:ras Proteins; D001920:Bradykinin
keywords: Bradykinin; Endothelial cell migration; Nitric oxide; Phosphatidyl-inositol-3-kinase; Rac1; Ras; S-nitrosylation