A Three-Stemmed mRNA Pseudoknot in the SARS Coronavirus Frameshift Signal.docx
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A Three-Stemmed mRNA Pseudoknot in the SARS Coronavirus Frameshift Signal.docx
AThree-StemmedmRNAPseudoknotintheSARSCoronavirusFrameshiftSignaloOpenaccess,freelyavaiIableonlineP1.SBIO1.OGYAThree-StemmedmRNAPseUdoknotintheSARSCOrOnaVirUSFranleShiftSignalEwanP.Plant1,GabrielaC.Perez-Alvarado2,Jonathan1.Jacobs1,BaniMukhopadhyay1,MirkoHennig2,JonathanD.Dinman1*1DepartmentofCe11BiologyandMo1ecu1arGenetics,UniversityofMaryland,CollegePark,Maryland,UnitedStatesofAmerica,2Departmentof-MolecIarBiologyandtKeSkaggslnstitUteforChemiCalBiology,ThescrippsResearchInstitute,1.aJolla,California,UnitedStatesofAmericaWiderangeofRNAvirusesuseprogrammedIribosoma1frameshiftIngfortheproductionofviralfusionproteins.InspectionoftheoverlapregionsbetweenORElaandORBlboftheSARS-CoVgenomerevealedthat,similartoallcoronaviruses,aprogrammed1ribosomalframeshiftcouldbeusedbythevirustoproduceafusionprotein.ComputationalanalysesoftheframeshiftsignalpredictedthepresenceofanmRNApseudoknotcontainingthreedouble-strandedRNAstemstructuresratherthantwo.BhylogeneticanalysesshowedthemedpseudoknotsintheframcshiftsignasofalIothercoronavirusesintHeGenBankdatabase.Thoughthepresenceofthethree-stemmedStructurcissupportedbynucleasemappingandtwo-dimensionalnuclearmagneticresonancestudies,ourfindingssuggesIlhatinteracIionsbctweenthestemstructuresmayresultJnlocaldistortionsinconservationofpotentialthree-stemtheA-formRNA.Theseclistortionsareparticularlycvidcntinthcvicinityofpredictedbu1gesinstems2and3.InvitroandinvivoframeshiftingassaysshowedthattheSARS-CoVframeshiftsignalisfunctionallysimiIartootherviralframeshiflsignals:ItpromotesefficientframeshiFtinginalIofthestandardassaysystems,anditissensitivetoadrugandagcneIicmutationthatareknowntoaffectframcshiftingefficiencyofayeastvirus.MutagenesiSstudiesrevea1thatboththespecificsequcncesandstructuresofstc11s2and3areimportantforeffIcientframeshifting.WehavemedidentifiedanewRNAstruetura1motifthatiscapab1eofpromo1.ingefficientprogrammedribosomalframeshifting.ThehighdegreeofconscrvatiOnOflhree-StCnlmRNpseudoknotstructu.
