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1、VirusResearch105(2004)121125CharacterizationofproteinproteininteractionsbetweenthenucleocapsidproteinandmembraneproteinoftheSARScoronavirusRuntaoHea,b,?,AndrewLeesona,MelissaBallantinea,AntonAndonova,b,LindsayBakera,FrederickDobiea,YanLia,b,NathalieBastiena,b,HeinzFeldmanna,b,UteStr
2、ochera,StevenTheriaulta,ToddCuttsa,JingxinCaoa,b,TimothyF.Bootha,b,FrankA.Plummera,b,ShaunTylera,XuguangLicaNationalMicrobiologyLaboratory,HealthCanada,1015ArlingtonSt.,Winnipeg,Man.,CanadaR3E3R2bDepartmentofMedicalMicrobiology,SchoolofMedicine,UniversityofManitoba,Winnipeg,Man.,Can
3、adaR3T2N2cCentreforBiologicsResearch,BiologicsandGeneticTherapiesDirectorate,HealthCanada,Tunney’sPasture,Ottawa,Ont.CanadaK1A0K9Received23February2004;receivedinrevisedform7May2004;accepted10May2004AbstractThehumancoronavirus,associatedwithsevereacuterespiratorysyndrome(SARS-CoV),w
4、asidenti?edandmolecularlycharacterizedin2003.Sequenceanalysisofthevirusindicatesthatthereisonly20%aminoacid(aa)identitywithknowncoronaviruses.Previousstudiesindicatethatproteinproteininteractionsamongstvariouscoronavirusproteinsarecriticalforviralassembly.Yet,littlesequencehomologyb
5、etweenthenewlyidenti?edSARS-CoVandthosepreviouslystudiedcoronavirusessuggeststhatdeterminationofproteinproteininteractionandidenti?cationofaminoacidsequences,responsibleforsuchinteractioninSARS-CoV,arenecessaryfortheelucidationofthemolecularmechanismofSARS-CoVreplicationandrationali
6、zationofanti-SARStherapeuticintervention.Inthisstudy,weemployedmammaliantwo-hybridsystemtoinvestigatepossibleinteractionsbetweenSARS-CoVnucleocapsid(N)andthemembrane(M)proteins.WefoundthatinteractionoftheNandMproteinstakesplaceinvivoandidenti?edthatastretchofaminoacids(168208)intheN
7、proteinmaybecriticalforsuchproteinproteininteractions.Importantly,thesameregionhasbeenfoundtoberequiredformultimerizationoftheNprotein(Heetal.,2004)suggestingthisregionmaybecrucialinmaintainingcorrectconformationoftheNproteinforself-interactionandinteractionwiththeMprotein.?2004Else
8、vierB.V.Allrightsre
