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1、VIROLOGY176,296-301(1990)DeducedSequenceoftheBovineCoronavirusSpikeProteinandIdentificationoftheInternalProteolyticCleavageSiteSUSHMAABRAHAM,THOMASE.KIENZLE,WILLIAMLAPPS,~ANDDAVIDA.BRIAN~DepartmentofMicrobiology,TheUniversityofTennessee,Knoxville,Tennessee37996-0845ReceivedOctober13,1
2、989;acceptedjanuary17,1990Thesequenceofthespike(alsocalledpeplomerorE2)proteingeneoftheMebusstrainofbovinecoronavirus(BCV)wasobtainedfromcDNAclonesofgenomicRNA.Thegenesequencepredictsa150,825molwtapoproteinof1363aminoacidshavinganN-terminalhydrophobicsignalsequenceof17aminoacids,19pot
3、entialN-linkedglycosylationsites,ahydrophobicanchorsequenceofapproximately17aminoacidsneartheCterminus,andahydrophiliccysteine-richCterminusof35aminoacids.AninternalLys-Arg-Arg-Ser-Arg-Argsequencepredictsaproteasecleavagesitebetweenaminoacids768and769thatwouldseparatetheSapoproteinint
4、oSlandS2segmentsof85690and65153molwt,respectively.Aminoterminalaminoacidsequencingofthevirion-derivedgplO0spikesubunitconfirmedthelocationofthepredictedcleavagesite,andestablishedthatgpl20andgplO0aretheglycosylatedvirionformsoftheSlandS2subunits,respectively.Sequencecomparisonsbetween
5、BCVandtheantigenicallyrelatedmousehepatitiscoronavirusrevealedmoresequencedivergenceintheputativeknobregionofthespikeprotein(Sl)thaninthestemregion(S2).0i990AcademicPress,Inc.Thebovinecoronavirus(BCV)isanimportantcausebethemajorstructurebywhichcoronavirusesattachofneonatalcalfdiarrhea
6、(14,26)andmayalsobethetocellsandinitiateinfection(reviewedbySpaaneta/.causeofwinterdysenteryinadultcattle(30).The(34)).TheHEprotein,however,mayalsobindtocellsmechanismsbywhichBCVcausesdiseaseandpersis-toinitiateinfection,andforBCV,therelativeimpor-tentinfectionarenotunderstood,norarec
7、urrentvac-tanceofthesetwoproteinsininitiatinginfectionisnotcinesuniversallyregardedaseffective.Towardtheseknown.BothSandHEareprobablyimportantininduc-ends,wehavebegunadetailedstudyoftheBCVpro-ingimmunitysinceantibodiestoeachareknowntoteinandgenomestructure.neutralizevirusinfectivityin
8、cellc
