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1、J.gen.ViroL(1985L66,593-596.PrintedinGreatBritain593Keywords:eoronavirusMHV/sequence/non-structuralprotein/hydrophobicityCodingSequenceofCoronavirusMHV-JHMmRNA4ByM.A.SKINNER*tANDS.G.SIDDELLInstituteofVirology,VersbacherStrasse7,8700Wi~rzburg,FederalRepublicofGermany(Accepted20N
2、ovember1984)SUMMARYAcodingsequenceatthe5'endofmRNA4ofthecoronavirusMHV-JHMwasdeterminedbyM13/chain-terminatorsequencingofclonedcDNA.Anopenreadingframeof417baseswiththepotentialtoencodeapolypeptideofmol.wt.15200(139residues)wasidentified.The3'endoftheopenreadingframeoverlappedby
3、16basesthestartofanopenreadingframefoundinmRNA5.ThetranslationproductofmRNA4waspredictedtobeabasicpolypeptiderichinthreonine.Ithadalargehydrophobicregionneartheaminoterminusandabasiccarboxyterminus.Anintracellular,virus-specificpolypeptide,whichhasbeenpreviouslydescribedashavin
4、gamol.wt.of14000to14500hasthesizeandchargecharacteristicsofsuchatranslationproduct.Murinehepatitisvirus(MHV)isamemberoftheCoronaviridaewhicharecytoplasmic,enveloped,RNAviruses,Themolecularbiologyofthisvirusgrouphasbeenreviewedrecently(Siddelletal.,1983).Briefly,incellsinfectedb
5、yMHV,inadditiontothegenomicRNA(tool.wt.6.0x106),sixsubgenomicmRNAswithmol.wt.rangingfrom0.6x106to3.7×106areproduced.Thegenomic-sizeRNAwhichisinfectiousistermedmRNA1andthesmallestsubgenomicmRNAismRNA7.TheseRNAspeciesformanestedset,eachRNAhaving3'sequencesincommonwithallsmallerRN
6、As.Atthe5'end,eachmRNAbearsacommonleaderofabout70basesderivedfromthe5'endofthegenome(Laietal.,1983;Spaanetal.,1983;Skinner&Siddell,1983;Armstrongetal.,1984a).Translationstudiesinvitroandinoocyteshaveshownthattheprimarytranslationproductsthatgiverisetothethreevirionstructuralpro
7、teins,peplomer(150K),membrane(26K)andnucleocapsid(50K),areproducedfrommRNAs3,6and7,respectively(Rottieretal.,1981;Siddelletal.,1981;Leibowitzetal.,1982).Thesizeofthesepolypeptidescorrespondswellwiththesizeofthe'unique'RNAofeachmessage,i.e.thatportionnotfoundinthenextsmallestmRN
8、A.SequencingofmRNA7(Skinner&Siddell,1983)andofmRNA6(Ar
