Analysis of Endocytic Pathways in Drosophila Cells Reveals a Conserved Role for GBF1 in Internalization via GEECs 英文参考文献.docVIP

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Analysis of Endocytic Pathways in Drosophila Cells Reveals a Conserved Role for GBF1 in Internalization via GEECs 英文参考文献.doc

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Analysis of Endocytic Pathways in Drosophila Cells Reveals a Conserved Role for GBF1 in Internalization via GEECs 英文参考文献

AnalysisofEndocyticPathwaysinDrosophilaCells RevealsaConservedRoleforGBF1inInternalizationvia GEECs GaganD.Gupta¤*,SwethaM.G.,SudhaKumari,RamyaLakshminarayan,GautamDey,SatyajitMayor* NationalCentreforBiologicalSciences,Bangalore,India Abstract In mammalian cells, endocytosis of the fluid phase and glycosylphosphatidylinositol-anchored proteins (GPI-APs) forms GEECs (GPI-AP enriched early endosomal compartments) via an Arf1- and Cdc42-mediated, dynamin independent mechanism.Hereweusefourdifferentfluorescentlylabeledprobesandseveralmarkersincombinationwithquantitative kinetic assays, RNA interference and high resolution imaging to delineate major endocytic routes in Drosophila cultured cells. We find that the hallmarks of the pinocytic GEEC pathway are conserved in Drosophila and identify garz, the fly orthologoftheGTPexchangefactorGBF1,asanovelcomponentofthispathway.LiveconfocalandTIRFimagingreveals thatafractionofGBF1GFPdynamicallyassociateswithABDRFP(asensorforactivatedArf1presentonnascentpinosomes). Correspondingly,aGTPexchangemutantofGBF1hasalteredABDRFPlocalizationintheevanescentfieldandisimpaired influidphaseuptake.Furthermore,GBF1activationisrequiredfortheGEECpathwayeveninthepresenceofBrefeldinA, implyingthat,likeArf1,ithasaroleinendocytosisthatisseparablefromitsroleinsecretion. Citation:GuptaGD,MGS,KumariS,LakshminarayanR,DeyG,etal.(2009)AnalysisofEndocyticPathwaysinDrosophilaCellsRevealsaConservedRoleforGBF1 inInternalizationviaGEECs.PLoSONE4(8):e6768.doi:10.1371/journal.pone.0006768 Editor:JeanGruenberg,UniversityofGeveva,Switzerland ReceivedMay14,2009;AcceptedJuly28,2009;PublishedAugust26,2009 Copyright: ? 2009 Gupta et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwasfundedinpartbyagrantfromtheNanotechnologymission[GovernmentofIndia,(GOI)].G.D.Gwassupportedbyfe