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1、VIROLOGY114.39-51(1981)TheVirus-SpecificIntracellularRNASpeciesofTwoMurineCoronaviruses:MHV-A59andMHV-JHMJULIANL.LEIBOWITZ,2KIRKC.WILHELMSEN;ANDCLIFFORDW.BOND4DepartmentofPathology,UniversityofCalifornia,SanDiego,SchoolofMedicine,LaJolla,California92093ReceivedFebruary20,1981;acceptedM
2、ay3,1981Sevenvirus-specific,polyadenylatedRNAspecieshavebeenidentifiedinmousecellsinfectedwiththemurinecoronavirusesMHV-A59(A59V)orMHV-JHM(JHMV).MHV-infected17CL.lcellswerelabeledwith[zrP]orthophosphateinthepresenceofactino-mycinDandtheeytoplasmicRNAwasextractedandanalyzedbyagarosegele
3、lectro-phoresis.TheseRNAspeciesrangeinsizefrom6.3X106to6.1Xlo6daltons.TheA59VandJHMV-specificRNAshaveidenticalmolecularweightsandcomigrateinagarosegels.ThelargestintracellularRNAspeciesisidenticaltoRNAisolatedfrompurifiedvirions,asdeterminedbyagarosegelelectrophoresisandoligonucleotide
4、fingerprintstudiesofribonucleaseT1digests.OligonucleotidefingerprintsofthesixsubgenomicRNAsshowthatthesequencestheycontainarepresentinvirionRNA,confirmingtheirvirus-specificnature.ThefingerprintingstudiesalsodemonstratethatthesixsubgenomicRNAspeciesmakeupanestedset.Thesequencespresenti
5、neachRNAspeciesarealsopresentinalllargerRNAspecies.TheselargerRNAsalsocontainadditionalsequencesconsistentwiththeirgreatersize.ThesubgenomicRNAsfulfullmanyofthecriteriaformRNAs.PossiblemechanismsforgeneratingtheseRNAsarediscussed.INTRODUCTIONbousprojectionsontheirsurface,budthroughthee
6、ndoplasmicreticulumratherCoronavirusesarewidespreadinnaturethantheplasmamembrane,andcontainandhavebeenassociatedwithseveraldis-RNA(Tyrelletal.,1978).Severalstudieseasesininfectedhosts(RobbandBond,ofcoronavirusgenomicRNAhavebeen1979a).Coronavirusesaredefinedaspleo-reported(Lomneczi,1977
7、;Lomnecziandmorphicenvelopedparticlesabout100nmKennedy,1977;Yogoetal.,1977;Laiandindiameterwhichhavecharacteristicbul-Stohlman,1978;MacnaughtonandMadge,1978;Schochetmanetal.,1977;TannockandHierholzer,1978;GuyandBrian,1979;iThisworkwassupportedinpartbyNIHGrantsNS07078,NS13898,andNS152
