Rotavirus Infection of Cells in Culture Induces Activation of RhoA and Changes in the Actin and Tubulin Cytoskeleton 英文参考文献.docVIP

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Rotavirus Infection of Cells in Culture Induces Activation of RhoA and Changes in the Actin and Tubulin Cytoskeleton 英文参考文献.doc

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Rotavirus Infection of Cells in Culture Induces Activation of RhoA and Changes in the Actin and Tubulin Cytoskeleton 英文参考文献

RotavirusInfectionofCellsinCultureInducesActivation ofRhoAandChangesintheActinandTubulin Cytoskeleton JoseLuisZambrano1*,OrlandoSorondo1,2,AnaAlcala1,EsmeraldaVizzi1,YuleimaDiaz3 ,Marie ChristineRuiz4,FabianMichelangeli4,FerdinandoLiprandi1,JuanE.Ludert5* 1InstitutoVenezolanodeInvestigacionesCient?′ficas(IVIC),CMBC.Caracas,Venezuela,2EscueladeBiolog?′a,UniversidadCentraldeVenezuela(UCV),Caracas,Venezuela, 3University of Bergen Thorm?hlensgate 55, Bergen, Norway, 4Instituto Venezolano de Investigaciones Cient?′ficas (IVIC), CBB. Caracas, Venezuela, 5Centro de Investigacio′nyEstudiosAvanzadosdelInstitutoPolite′cnicoNacional(CINVESTAV-IPN),CiudaddeMe′xico,Me′xico Abstract Rotavirusinfectioninducesanincreasein[Ca2+]cyto,whichinturnmayaffectthedistributionofthecytoskeletonproteinsin the infected cell. Changes in microfilaments, including the formation of stress fibers, were observed starting at 0.5h.p.i. using fluorescent phalloidin. Western blot analysis indicated that RhoA is activated between 0.5 and 1h.p.i. Neither the phosphorylationofRhoAnortheformationofstressfiberswereobservedincellsinfectedwithvirionspre-treatedwithan anti-VP5*non-neutralizingmAb,suggestingthatRhoAactivationisstimulatedbytheinteractionoftheviruswithintegrins formingthecellreceptorcomplex.Inaddition,thestructureofthetubulincytoskeletonwasalsostudied.Alterationsofthe microtubules were evident starting at 3h.p.i. and by 7h.p.i. when microtubules were markedly displaced toward the peripheryofthecellcytoplasm.Loadingofrotavirus-infectedcellswitheitheraCa2+chelator(BAPTA)ortransfectionwith siRNAstosilenceNSP4,reversedthechangesobservedinboththemicrofilamentsandmicrotubulesdistribution,butnot theappearanceofstressfibers.Theseresultsindicatethatalterationsinthedistributionofactinmicrofilamentsareinitiated earlyduringinfectionbytheactivationofRhoA,andthatlatterchangesintheCa2+homeostasispromotedbyNSP4during infectionmayberesponsibleforotheralterationsintheactinandtubulincytoskeleton. Citation:ZambranoJ