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1、VIROLOGY165,367-376(1988)TheAmino-TerminalSignalPeptideonthePorcineTransmissibleGastroenteritisCoronavirusMatrixProteinIsNotanAbsoluteRequirementforMembraneTranslocationandGlycosylationPAULA.KAPKE,*FRANKY.T.TUNG,tBRENDAG.HOGUE,+’DAVIDA.BRlAN,tv2ROGERD.WOODS,*ANDRONALDWESLEY*lU,
2、S.DepartmentofAgriculture,AgriculturalResearchService,NationalAnimalDiseaseCenter,P.O.Box70Ames,lowa50070,andtDepartmentofMicrobiology,TheUniversityofTennessee,Knoxville.Tennessee37996.0845ReceivedDecember18,1987;acceptedApril6,1988cDNAclonesmappingwithinthefirst2601basesofthe3
3、’endoftheporcinetransmissiblegastroenteritiscorona-virus(TGEV)genomeweresequencedbythemethodofMaxamandGilbertandanopenreadingframeyieldingaproteinhavingpropertiesofthematrix(MorEl)proteinwasidentified.Itispositionedatthe5’sideofthenucleocap-sid(N)genefromwhichitisseparatedbyani
4、ntergenicstretchof12bases.ThededucedMproteincomprises262aminoacids,hasamolecularweightof29,544,ismoderatelyhydrophobic,andhasanetchargeoft7atneutralpH.Thirty-fourpercentofitsaminoacidsequenceishomologouswiththeMproteinofthebovinecoronavirus(BCV),32%withthatofthemousehepatitisco
5、ronavirus(MHV),and19%withthatoftheavianinfectiousbronchitiscoronavirus(IBV).JudgingfromalignmentwiththeBCV,MI-IV,andIBVMproteins,theaminoterminusoftheTGEVMproteinextends54aminoacidsfromthevirionenvelopewhichcompareswithonly28forBCV,26forMHV,and21forIBV.Elevenofthesixteenamino-t
6、erminalaminoacidsarehydrophobicandthepositionsofchargedaminoacidsaroundthissequencesuggestthatthefirst16aminoacidscompriseapotentiallycleavablesignalpeptideformembraneinsertion.AsimilarsequenceisnotfoundintheMproteinsofBCV,MHV,orIBV.WhenmRNAfrominfectedcells,orRNApreparedbyinvi
7、trotranscriptionofthereconstructedMgene,wastranslatedinvitrointhepresenceofmicrosomes,theMproteinbecametranslocatedandglycosylated.Whenaproteinwithouttheamino-terminalsignalpeptidewasmadebytranslatingatruncatedversionoftheMgenetranscript,sometranslocationandglycosylationalsoocc
8、urredsuggestingthattheamino-terminalsignalpeptideonthe
