Rgnef (p190RhoGEF) Knockout Inhibits RhoA Activity, Focal Adhesion Establishment, and Cell Motility Downstream of Integrins 英文参考文献.docVIP

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Rgnef (p190RhoGEF) Knockout Inhibits RhoA Activity, Focal Adhesion Establishment, and Cell Motility Downstream of Integrins 英文参考文献.doc

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Rgnef (p190RhoGEF) Knockout Inhibits RhoA Activity, Focal Adhesion Establishment, and Cell Motility Downstream of Integrins 英文参考文献

Rgnef(p190RhoGEF)KnockoutInhibitsRhoAActivity, FocalAdhesionEstablishment,andCellMotility DownstreamofIntegrins NicholL.G.Miller,ChristineLawson,XiaoLeiChen,Ssang-TaekLim¤,DavidD.Schlaepfer* MooresCancerCenter,UniversityofCaliforniaSanDiego,LaJolla,California,UnitedStatesofAmerica Abstract Background:Cellmigrationisahighlyregulatedprocessthatinvolvestheformationandturnoverofcell-matrixcontact sitestermedfocaladhesions.Rho-familyGTPasesaremolecularswitchesthatregulateactinandfocaladhesiondynamicsin cells.Guaninenucleotideexchangefactors(GEFs)activateRho-familyGTPases.Rgnef(p190RhoGEF)isaubiquitous190kDa GEFimplicatedinthecontrolofcoloncarcinomaandfibroblastcellmotility. Principal Findings: Rgnef exon 24 floxed mice (Rgnefflox) were created and crossed with cytomegalovirus (CMV)-driven Cre recombinasetransgenicmicetoinactivateRgnefexpressioninalltissuesduringearlydevelopment.HeterozygousRgnefWT/flox (Cre+)crossesyieldednormalMendelianratiosatembryonicday13.5,butRgnefflox/flox(Cre )micenumbersat3weeksofagewere + (Cre ) (Rgnef2/2) embryos and primary mouse embryo fibroblasts (MEFs) were + flox/flox significantly less than expected. Rgnef isolatedandverifiedtolackRgnefproteinexpression.Whencomparedtowildtype(WT)littermateMEFs,lossofRgnefsignificantly inhibited haptotaxis migration, wound closure motility, focal adhesion number, and RhoA GTPase activation after fibronectin- integrinstimulation.InWTMEFs,Rgnefactivationoccurswithin60minutesuponfibronectinplatingofcellsassociatedwithRhoA activation.Rgnef2/2MEFphenotypeswererescuedbyepitope-taggedRgnefre-expression. Conclusions:Rgnef2/2MEFphenotypeswereduetoRgneflossandsupportanessentialroleforRgnefinRhoAregulation downstreamofintegrinsincontrolofcellmigration. Citation:MillerNLG,LawsonC,ChenXL,LimS-T,SchlaepferDD(2012)Rgnef(p190RhoGEF)KnockoutInhibitsRhoAActivity,FocalAdhesionEstablishment,and CellMotilityDownstreamofIntegrins.PLoSONE7(5):e37830.doi:10.1371/journal.pone.0037830 Editor:MaddyParsons,King’sCollegeLondon,U