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1、VirusResearchELSEVIERVirusResearch39(1995)261-276Aconditional-lethalmurinecoronavirusmutantthatfailstoincorporatethespikeglycoproteinintoassembledvirionsCynthiaS.Ricarda,1,CheriA.Koetznerb,LawrenceS.Sturmanb,c,PaulS.Mastersb,c,.aDepartmentofMicrobiology,Immunology,andMolecularGenetic
2、s,AlbanyMedicalCollege,Albany,NY12208,USAbWadsworthCenterforLaboratoriesandResearch,NewYorkStateDepartmentofHealth,Albany,NY12201,USAcDepartmentofBiomedicalSciences,StateUniversityofNewYorkatAlbany,Albany,NY12237,USAReceived4August1995;revised11September1995;accepted12September1995Ab
3、stractThecoronavirusspikeglycoprotein(S)mediatesboththeattachmentofvirustothehostcellreceptorandmembranefusion.Wedescribeherethecharacterizationofatempera-ture-sensitivemutantofthecoronavirusmousehepatitisvirusA59(MHV-A59)havingmultipleSprotein-relateddefects.Themostremarkableofthese
4、wasthatthemutant,designatedAlbany18(Albl8),assembledvirionsdevoidoftheSglycoproteinatthenonpermissivetemperature.Albl8alsofailedtobringaboutsyncytiaformationincellsinfectedatthenonpermissivetemperature.Virionsofthemutantassembledatthepermis-sivetemperatureweremuchmorethermolabilethan
5、wildtype.Moreover,mutantSproteinthatwasincorporatedintovirionsatthepermissivetemperatureshowedenhancedpH-de-pendentthermolabilityinitsabilitytobindtotheMHVreceptor.Albl8wasfoundtohaveasinglepointmutationinSresultinginachangeofserine287toisoleucin¢,anditwasshownbyrevertantanalysisthat
6、thiswasthelesionresponsibleforthephenotypeofthemutant.Keywords:Coronavirus;Spikeglycoprotein;Temperature-sensitivemutant;Virusassembly1Presentaddress:DepartmentofBiochemistryandMolecularBiophysics,WashingtonUniversitySchoolofMedicine,Box8231,660SouthEuclidAvenue,St.Louis,MO63110,USA.
7、*Correspondingauthorataddressb.Tel.:+1(518)474-1283;Fax:+1(518)473-1326.0168-1702/95/$09.50©1995ElsevierScienceB.V.AllrightsreservedSSDI0168-1702(95)00100-X262C.S.Ricardetal./I4rusResearch39(1995)261-276I.IntroductionCoronavirusesareafamilyofenveloped,positive-strandRNAvirusescausing
8、anumberofdis
