冠狀病毒Coronavirusl論文-2004 The Severe Acute Respiratory Syndrome Coronavirus Nsp15 Protein Is an Endoribonuclease That Prefers Manganese as a.pdf

冠狀病毒Coronavirusl論文-2004 The Severe Acute Respiratory Syndrome Coronavirus Nsp15 Protein Is an Endoribonuclease That Prefers Manganese as a.pdf

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1、JOURNALOFVIROLOGY,Nov.2004,p.1221812224Vol.78,No.220022-538X/04/$08.000DOI:10.1128/JVI.78.22.1221812224.2004Copyright?2004,AmericanSocietyforMicrobiology.AllRightsReserved.TheSevereAcuteRespiratorySyndromeCoronavirusNsp15ProteinIsanEndoribonucleaseThatPrefersManganeseasaCofactorKanchanBhard

2、waj,LindaGuarino,andC.ChengKao*DepartmentofBiochemistryandBiophysics,TexasA&MUniversity,CollegeStation,TexasReceived23April2004/Accepted9July2004Nonstructuralprotein15(Nsp15)ofthesevereacuterespiratorysyndromecoronavirus(SARS-CoV)pro-ducedinEscherichiacolihasendoribonucleaseactivitythatprefe

3、rentiallycleaved5ofuridylatesofRNAs.Blockingeitherthe5or3terminusdidnotaffectcleavage.Double-andsingle-strandedRNAswereboth2substratesforNsp15butwithdifferentkineticsforcleavage.Mnat2to10mMwasneededforoptimal2endoribonucleaseactivity,butMgandseveralotherdivalentmetalswerecapableofsuppor

4、tingonlyalow2levelofactivity.ConcentrationsofMnneededforendoribonucleaseactivityinducedsigni?cantconforma-tionchange(s)intheprotein,asmeasuredbychangesintryptophan?uorescence.Asimilarendoribonucleo-lyticactivitywasdetectedfortheorthologousproteinfromanothercoronavirus,demonstratingthattheen

5、doribonucleaseactivityofNsp15maybecommontocoronaviruses.Thisworkpresentsaninitialbiochemicalcharacterizationofanovelcoronavirusendoribonuclease.Thesevereacuterespiratorysyndrome(SARS)epidemicofNsp13isahelicasewiththeabilitytounwindbothDNAand2002to2003eventuallyspreadfromChinato30countries,RN

6、Aduplexes.Nsp13isalsoreportedtohavedNTPaseandwithmorethan8,000con?rmedcasesandanoverallmortalityRNA5-triphosphataseactivities(6).rateofabout10%(31).Duetothelackofmorespeci?can-Ingeneral,thebiochemicalcharacterizationsofthecoro-tiviralagents,onetreatmentregimenusedconsistedofcorti-navirusrep

7、lication-associatedproteinsareatanearlystage.coidsteroidsandribavirin(15,16).Thisregimenwasofun-SnijderandcolleagueshaveproposedthatNsp15hassimilar-2knownef?cacyandhadseveresideeffects.Thus,thereisaitiestotheendoribonucleaseXendoU(25),aMn-depen-ne

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