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1、Proc.Nati.Acad.Sci.USAVol.84,pp.6567-6571,September1987MicrobiologyRNArecombinationofcoronaviruses:LocalizationofneutralizingepitopesandneuropathogenicdeterminantsonthecarboxylterminusofpeplomersSHINJIMAKINO,JOHN0.FLEMING,JAMESG.KECK,STEPHENA.STOHLMAN,ANDMI
2、CHAELM.C.LAIDepartmentsofMicrobiologyandNeurology,UniversityofSouthernCalifornia,SchoolofMedicine,LosAngeles,CA90033CommunicatedbyPeterK.Vogt,May11,1987(receivedforreviewFebruary2,1987)ABSTRACTMurinecoronavirusesundergoRNArecom-ofeachmRNAiscontainedentirely
3、withinthenextlargerbinationataveryhighfrequency.WehaveobtainedaseriesofmRNA(10).Onlythe5'-specificportions,whichdonotrecombinantvirusesusingneutralizingmonoclonalantibodiesoverlapwiththenextsmallermRNAs,aretranslated.Fur-inconjunctionwithtemperature-sensiti
4、vemarkers.Allofthethermore,eachmRNAcontainsaleadersequenceof'70recombinantsobtainedhaveacrossoverwithingeneC,whichnucleotides,whichisderivedfromthe5'endofthegenomeencodesthepeplomerproteinofthevirus.Thegeneticstruc-andjoinedtothemRNAsbyamechanismof"leader-p
5、rimedtureoftheserecombinantssuggeststhattheantigenicregionstranscription"(11,12).DespitethefactthatMHVgenomeisrecognizedbytheseneutralizingmonoclonalantibodiesareasingle,nonsegmentedRNA,ourlaboratoryhasdemon-localizedonthecarboxyl-terminalone-thirdofthepepl
6、omerstratedthatMHVcanundergoRNA-RNArecombinationatprotein.Sincethetwomonoclonalantibodiesusedarealsoanextremelyhighfrequency(13),reminiscentofthegeneticassociatedwiththecriticaldeterminantsofvirusneuropatho-reassortmentofviruseswithsegmentedRNAgenomes.Thisg
7、enicity,weconcludethatboththeneutralizingantibodyobservationsuggeststhatfragmentedRNAintermediatesbindingsitesanddeterminantsofpathogenicityarelocalizedatmightbegeneratedduringreplicationofMHVRNA,prob-thecarboxyl-terminalone-thirdofthepeplomer.Thevariationa
8、blybyamechanismofdiscontinuousandnonprocessiveofcrossoversitesindifferentrecombinantvirusesalsoallowedRNAsynthesis(13,14).TheavailabilityofRNArecombi-precisemappingofadditionalantigenicsites.RNArecombi
