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1、VIROLOGY175,548-555(1990)AClusteringofRNARecombinationSitesAdjacenttoaHypervariableRegionofthePeplomerGeneofMurineCoronavirusLISAR.BANNER,JAMESG.KECK,’ANDMICHAELM.C.LA12DepartmentofMicrobiology,UniversityofSouthernCaliforniaSchoolofMedicine,LosAngeles,California,90033ReceivedOctober12,1989;a
2、cceptedDecember12,1989CoronavirusesundergoRNArecombinationataveryhighfrequency.Tounderstandthemechanismofrecombina-tioninmurinecoronavirus,wehaveperformedRNAsequencingofviralgenomicRNAtodeterminetheprecisesitesofrecombinationinaseriesofrecombinantswhichhavecrossoverswithinthegeneencodingthep
3、eplomerprotein.Wefoundthatalloftherecombinationsitesareclusteredwithinaregionof278nucleotidesinthe5’-halfofthegene.Thisregioninwhichallofthecrossoversoccurredrepresentsasmallfractionofthedistancebetweenthetwoselectionmarkersusedfortheisolationoftheserecombinantviruses.Thisresultsuggeststhatt
4、hisregionmaybeapreferredsiteforRNArecombination.Thecrossoversitesarelocatedwithinandimmediatelyadjacenttoahypervariableareaofthegene.Thisareahasundergonedeletionsofvaryingsizesinseveralvirusstrainswhichhavebeenpassagedeitherinvivaorinvitro.TheseresultssuggestthatasimilarRNAstructuremaybeinvo
5、lvedintheoccurrenceofbothrecombinationanddeletionevents.o199OAcademicPWSS,I~C.INTRODUCTIONgp65)containsanesteraseactivitybutispresentonlyincertainstrainsofMHV(LaiandStohlman,1981;Sid-Mousehepatitisvirus(MHV),theprototypicmemberdell,1982;Makinoetal.,1983;MakinoandLai,1989;oftheCoronaviridae,i
6、sanenvelopedviruswhichcon-Shieheta/.,1989).TheMprotein(formerlynamedEl)tainsnonsegmentedpositive-sensedRNAof32kbinismostlyembeddedwithintheviralenvelope(Arm-length(LaiandStohlman,1978;Pachucketal.,1989;strongeta/.,1984;Rottiereta/.,1984,1986);however,Bakereta/.,1989).Thevirusparticleiscompos
7、edofmonoclonalantibodiesagainstMproteincanalsoalterthreeenvelopeproteins(S,M,andHE)andanucleo-thepathogenicityofthevirus(Flemingeta/.,1989).capsidprotein(N).TheSprotein(Spikeprotein,for-AlloftheviralproteinsaretranslatedfromseparatemerlynamedE2)for
