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1、VIROLOGY173,683-691(1989)Biosynthesis,Structure,andBiologicalActivitiesofEnvelopeProteingp65ofMurineCoronavirusKYOKOYOKOMORI,NlCOL4LAMONICA,SHINJIMAKINO,CHIEN-KOUSHIEH,ANDMICHAELM.C.LAI’DepartmentofMicrobiology,UniversityofSouthernCalifornia,SchoolofMedicine,LosAngeles,
2、California90033ReceivedJune21I1989;acceptedAugust25,1989Wehavepreviouslyshownthatgp65(E3)isavirionstructuralproteinwhichvarieswidelyinquantityamongdifferentstrainsofmousehepatitisvirus(MHV).Inthisstudy,thebiosyntheticpathwayandpossiblebiologicalactivitiesofthisproteinwe
3、reexamined.Theglycosylationofgp65invirus-infectedcellswasinhibitedbytunicamycinbutnotbymonen-sin,suggestingthatitcontainsan/V-glycosidiclinkage.GlycosylationiscotranslationalandappearstobecompletebeforetheglycoproteinreachestheGolgicomplex.Pulse-chaseexperimentsshowedth
4、atthisproteindecreasedinsizeafter30minofchase,suggestingthatthecarbohydratechainsofgp65undergotrimmingduringitstransportacrosstheGolgi.Thisinterpretationissupportedbytheendoglycosidasetreatmentofgp65,whichshowedthatthepeptidebackboneofgp65didnotdecreaseinsizeafterpulse-
5、chaseperiods.ThismaturationpathwayisdistinctfromthatoftheElorE2glycoproteins.Partialendoglycosidasetreatmentindicatedthatgp65contains9to10carbohydratesidechains;thus,almostallofthepotentialglycosylationsitesofgp65wereglycosylated.Invitrotranslationstudiescoupledwithprot
6、easedigestionsuggestthatgp65isanintegralmembraneprotein.Thepresenceofgp65inthevirioniscorrelatedwiththepresenceofanacetylesteraseactivity.Nohemagglutininactivitywasdetected.o1989AcademicPress,Inc.INTRODUCTIONprotein,partofwhichisexposedontheviralsurfaceofthisvirus(Makin
7、oetal.,1983).Mousehepatitisvirus(MHV),amemberoftheCoro-ThestructuralproteinsofMHVaretranslatedfromnaviridaefamily,isanenvelopedviruswithapositive-virus-specificmonocistronicmRNAsinthevirus-in-sensedRNAgenomeofmorethan6Xlo6molecularfectedcells(Rottieretal.,1981;Leibowitz
8、etal.,1982).weight(LaiandStohlman,1978;Wegeeta/.,1978).ThesemRNAshavea3’-coterminalnested-setstruc-Thevirionpa
