A Three-Stemmed mRNA Pseudoknot in the SARS Coronavirus Frameshift Signal.docx

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1、AThree-StemmedmRNAPseudoknotintheSARSCoronavirusFrameshiftSignaloOpenaccess,freelyavaiIableonlineP1.SBIO1.OGYAThree-StemmedmRNAPseUdoknotintheSARSCOrOnaVirUSFranleShiftSignalEwanP.Plant1,GabrielaC.Perez-Alvarado2,Jonathan1.Jacobs1,BaniMukhopadhyay1,MirkoHennig2,JonathanD.Dinman1*1DepartmentofCe11Bio

2、logyandMo1ecu1arGenetics,UniversityofMaryland,CollegePark,Maryland,UnitedStatesofAmerica,2Departmentof-MolecIarBiologyandtKeSkaggslnstitUteforChemiCalBiology,ThescrippsResearchInstitute,1.aJolla,California,UnitedStatesofAmericaWiderangeofRNAvirusesuseprogrammedIribosoma1frameshiftIngfortheproduction

3、ofviralfusionproteins.InspectionoftheoverlapregionsbetweenORElaandORBlboftheSARS-CoVgenomerevealedthat,similartoallcoronaviruses,aprogrammed1ribosomalframeshiftcouldbeusedbythevirustoproduceafusionprotein.ComputationalanalysesoftheframeshiftsignalpredictedthepresenceofanmRNApseudoknotcontainingthree

4、double-strandedRNAstemstructuresratherthantwo.BhylogeneticanalysesshowedthemedpseudoknotsintheframcshiftsignasofalIothercoronavirusesintHeGenBankdatabase.Thoughthepresenceofthethree-stemmedStructurcissupportedbynucleasemappingandtwo-dimensionalnuclearmagneticresonancestudies,ourfindingssuggesIlhatin

5、teracIionsbctweenthestemstructuresmayresultJnlocaldistortionsinconservationofpotentialthree-stemtheA-formRNA.Theseclistortionsareparticularlycvidcntinthcvicinityofpredictedbu1gesinstems2and3.InvitroandinvivoframeshiftingassaysshowedthattheSARS-CoVframeshiftsignalisfunctionallysimiIartootherviralfram

6、eshiflsignals:ItpromotesefficientframeshiFtinginalIofthestandardassaysystems,anditissensitivetoadrugandagcneIicmutationthatareknowntoaffectframcshiftingefficiencyofayeastvirus.MutagenesiSstudiesrevea1thatboththespecificsequcncesandstructuresofstc11s2and3areimportantforeffIcientframeshifting.WehavemedidentifiedanewRNAstruetura1motifthatiscapab1eofpromo1.ingefficientprogrammedribosomalframeshifting.ThehighdegreeofconscrvatiOnOflhree-StCnlmRNpseudoknotstructu.