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1、Proc.Nati.Acad.Sci.USAVol.83,pp.4204-4208,June1986BiochemistryLeadersequencesofmurinecoronavirusmRNAscanbefreelyreassorted:EvidencefortheroleoffreeleaderRNAintranscription(RNaseH/reassortmentofleaderRNAsequences/transacting)SHINJIMAKINO,STEPHENA.STOHLM
2、AN,ANDMICHAELM.C.LAIDepartmentsofMicrobiologyandNeurology,UniversityofSouthernCaliforniaSchoolofMedicine,LosAngeles,CA90033CommunicatedbyPeterK.Vogt,February18,1986ABSTRACTMousehepatitisvirus(MHV),whichrepli-transcriptioncomplex(12).Wehavealsoisolateda
3、temper-catesincytoplasmofinfectedcells,containsanidenticalleaderature-sensitivemutantthatsynthesizesonlysmallleaderRNAsequenceatthe5'endofeachofthevirus-specificRNAs,butnotmRNAs,atnonpermissivetemperature(12).mRNAs.Previousstudiessuggestedthatthesynthe
4、sisoftheseThesedatasuggestthatMHVmRNAsaresynthesizedmRNAsdoesnotinvolveconventionalRNAsplicingandmaydiscontinuouslyandmayinvolveafreeleaderRNAforinsteadrequireprimingbyafreeleaderRNA.Inthiscom-transcription.Thereis,however,nodirectevidencethatmunicatio
5、n,wedemonstratethat,duringamixedinfectionthesefreeleader-sequence-containingsmallRNAspecieswithtwodifferentMHVs,theleaderRNAsequencesfromonedetectedareutilizedforMHVRNAtranscriptionandarenotviruscouldbedetectedonthemRNAsofthecoinfectingvirusmerelyabort
6、ivetranscriptionproducts.atahighfrequency,asiftheleadersequenceandmRNAswereInthisinvestigation,weexaminedwhetherthefreeleaderjoinedtogetherfromtworandomlysegregatingRNAsegments.RNAspeciesaretrueintermediatesofMHVRNAtranscrip-ThisrmdingdemonstratesthatM
7、HVmRNAtranscriptiontionbystudyingtheleadersequencesofmRNAssynthesizedutilizesindependentlytranscribedleaderRNAspeciesthatincellscoinfectedwithtwodifferentstrainsofMHV.IfMHVpossessthetrans-actingproperty.ThisstudythusprovidestranscriptionusesfreeleaderR
8、NAspeciesfromacytoplas-furtherevidenceinsupportoftheuniquemodelof"leader-micpoolasaprimer,theleadersequencescanbeexpectedprimedtranscription"forcoronavirusmRNAsynthesis.tobereassortedfreelybetweenthemRNAsofthetwodifferentMHVs.Wehaveinde
