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1、JOURNALOFVIROLOGY,Oct.2007,p.1152011525Vol.81,No.200022-538X/07/$08.000doi:10.1128/JVI.01308-07Copyright?2007,AmericanSocietyforMicrobiology.AllRightsReserved.NOTESEnhancementofMurineCoronavirusReplicationbySevereAcuteRespiratorySyndromeCoronavirusProtein6RequirestheN-TerminalHydrophobicRegi
2、onbutNotC-TerminalSortingMotifs1223JasonNetland,DebraFerraro,LeciaPewe,HeidiOlivares,31,2ThomasGallagher,andStanleyPerlman*12InterdisciplinaryPrograminImmunologyandDepartmentofMicrobiology,UniversityofIowa,IowaCity,Iowa,and3DepartmentofMicrobiologyandImmunology,LoyolaUniversityMedicalCenter,
3、Maywood,IllinoisReceived14June2007/Accepted23July2007Severeacuterespiratorysyndromecoronavirusencodesseveralaccessoryproteinsofunknownfunction.Wepreviouslyshowedthatonesuchprotein,encodedbyORF6,enhancedthegrowthofmousehepatitisvirusintissueculturecellsandinmice.Protein6consistsofanN-terminalh
4、ydrophobicpeptideandaC-terminalregioncontainingintracellularproteinsortingmotifs.Herein,weshowthatmutationofthehydrophobicregionbutnotthesortingmotifsaffectedtheabilityofprotein6toenhancevirusgrowth.Collectively,theseresultssupportthenotionthatthe6proteininteractswithmembrane-boundviralreplic
5、ationorassemblymachinerytodirectlyenhancevirusreplicationandvirulenceinanimals.Theetiologicalagentofthesevereacuterespiratorysyn-amphipathicportionortheputativeproteinsortingmotifsindrome(SARS)wasidenti?edasacoronavirus(SARS-CoV).theCterminusareessentialforprotein6toin?uenceCoVInadditionaltos
6、tructuralproteins,SARS-CoVencodeseightinfections.accessoryproteins,whosefunctionsarenotwellde?ned(15).Asinourpreviousstudies,weusedJ2.2-v-1astheparentalPreviously,weintroducedeachoftheSARS-CoVaccessoryvirus(13).VariantsweregeneratedbyPCRusingprimersgenesintoanattenuatedstrainofmousehepatitisv
7、irusencodingthedesiredORF6mutations(primersequences(MHV),strainJHMJ2.2-v-1,(J2.2-v-1),byusingreversege-availableuponrequest).Inthe?rstsetofmutants,YSELandnetics(14).J2.2-v-1causesamildencephalitisfollowedbyadiacidicsortingmotifslocatednearthe
