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1、VIROLOGY116,8tid-a%4(1981)TunicamycinResistantGlycosylationofaCoronavirusGlycoprotein:DemonstrationofaNovelTypeofViralGlycoproteinKATHRYNV.HOLMES,**lELIZABETHW.DOLLER,*ANDLAWRENCES.STURMANt*DepartmenfofPathology,TheUn$brmedseruicesUniversityoftheHealthsciaces,Bethesda,Margl
2、and24IOl4and~DivisiaofLubmatoriasandResewch,NewYmkStateDepartmentofHealth,Albany,NewYork122y)lReceivedJuneS,1981;acceptedAugust3,1981Tunicamycinhasdifferenteffectsontheglycosylationofthetwoenvelopeglycoproteinsofmousehepatitisvirus(MHV),acoronavirus.Unlikeenvelopeglycoprote
3、insofotherviruses,thetransmembraneglycoproteinElisglycosylatednormallyinthepresenceoftunicamycin.ThissuggeststhatglycosylationofEldoesnotinvolvetransferofcoreoligosaccharidesfromdolicholpyrophosphateintermediatestoasparagineresidues,butmayoccurbyO-linkedglycosylationofserin
4、eorthreonineresidues.SynthesisofthepeplomericglycoproteinE2isnotreadilydetectableinthepresenceoftunicamycin.InhibitionofN-linkedglycosylationofE2bytunicamycineitherpreventssynthesisorfacilitatesdegradationoftheproteinmoietyofE2.Radiolabelingwithcarbohydrateprecursorsandbora
5、tegelelectrophoresisofglycopeptidesshowthatdifferentoligosac-charidesidechainsareattachedtoElandE2.Thetwocoronavirusenvelopeglycoproteinsthusappeartobeglycosylatedbydifferentmechanisms.Intunicamycin-treatedcells,noninfectiousvirionslackingpeplomersareformedatintracytoplasmi
6、cmembranesandreleasedfromthecells.ThesevirionscontainnormalamountsofnucleocapsidproteinandglycosylatedEl,butlackE2.ThusthetransmembraneglycoproteinElistheonlyviralglycoproteinrequiredfortheformationoftheviralenvelopeorforvirusmaturationandrelease.ThepeplomericglycoproteinE2
7、appearstoberequiredforattachmenttovirusreceptorsontheplasmamembrane.ThecoronavirusenvelopeenvelopeglycoproteinElappearstobeanoveltypeofviralglycoproteinwhichispost-translationallyglyco-sylatedbyatunicamycin-resistantprocessthatyieldsoligosaccharidesidechainsdif-ferentfromth
8、oseofN-linkedglycoproteins.ThesefindingssuggestthatElmaybeparticularlyusefulasamod
