Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells Toward the Identification of Budding Inhibitors英文文献资料.docVIP

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Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells Toward the Identification of Budding Inhibitors英文文献资料.doc

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Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells Toward the Identification of Budding Inhibitors英文文献资料

HindawiPublishingCorporation AdvancesinVirology Volume2011,ArticleID341816,10pages doi:10.1155/2011/341816 ResearchArticle BimolecularComplementationtoVisualize FilovirusVP40-HostComplexesinLiveMammalianCells: TowardtheIdenti?cationofBuddingInhibitors YuliangLiu, MichaelS.Lee, MarkA.Olson, andRonaldN.Harty 1 2 2 1 1 DepartmentofPathobiology,SchoolofVeterinaryMedicine,UniversityofPennsylvania,3800SpruceStreet,Philadelphia, PA19104,USA 2 DepartmentofCellBiologyandBiochemistry,U.S.ArmyMedicalResearchInstituteofInfectiousDiseases,1425PorterStreet, Frederick,MD21702,USA CorrespondenceshouldbeaddressedtoRonaldN.Harty,rharty@ Received8July2011;Accepted9August2011 AcademicEditor:AnthonyP.Schmitt Copyright?2011YuliangLiuetal.ThisisanopenaccessarticledistributedundertheCreativeCommonsAttributionLicense, whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperlycited. Virus-hostinteractionsplaykeyrolesinpromotinge?cientegressofmanyRNAviruses,includingEbolavirus(EBOVor“e”)and Marburgvirus(MARVor“m”).Late-(L-)domainsconservedinviralmatrixproteinsrecruitspeci?chostproteins,suchasTsg101 andNedd4,tofacilitatethebuddingprocess.Theseinteractionsserveasattractivetargetsforthedevelopmentofbroad-spectrum buddinginhibitors.Amajorgapstillexistsinourunderstandingofthemechanismof?lovirusbuddingduetothedi?cultyin detectingvirus-hostcomplexesandmappingtheirtra?ckingpatternsinthenaturalenvironmentofthecell.Toaddressthisgap, weusedabimolecularcomplementation(BiMC)approachtodetect,localize,andfollowthetra?ckingpatternsofeVP40-Tsg101 complexesinlivemammaliancells.Inaddition,weusedtheBiMCapproachalongwithaVLPbuddingassaytotestsmallmolecule inhibitorsidenti?edbyinsilicoscreeningfortheirabilitytoblockeVP40PTAP-mediatedinteractionswithTsg101andsubsequent buddingofeVP40VLPs.Wedemonstratedthepotentialbroadspectrumactivityofaleadcandidateinhibitorbydemonstrating itsabilitytoblockPTAP-dependentbindingofHIV-1