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1、ProteomicsClin.Appl.2011,5,460482467ThisarticlewasoriginallypublishedinProteomics2011,11,6480,DOI10.1002/pmic.201000309IdentificationofhostfactorsinvolvedincoronavirusreplicationbyquantitativeproteomicsanalysisMijkeW.Vogels,BasW.M.vanBalkom,DoraV.Kaloyanov
2、a,JosephJ.Batenburg,AlbertJ.Heck,J.BerndHelms,PeterJ.M.RottierandCornelisA.M.deHaanKeywords:Hostpathogeninteractions/Microbiology/Secretorypathway/SILACInthisstudy,weappliedaquantitativeproteomicapproach,basedonSILAC,toinvestigatetheinteractionsofcorona-v
3、iruseswiththesecretorypathwayofthehostcell,withtheaimtoidentifyhostfactorsinvolvedincoronavirusreplication.ComparisonoftheproteinprofilesofGolgi-enrichedfractionsofcellsthatwereeithermockinfectedorinfectedwithmousehepatitisvirusrevealedthesignificantdepleti
4、onorenrichmentof116proteins.Althoughriboso-mal/nucleicacidbindingproteinswereenrichedintheGolgi-fractionsofmousehepatitisvirus-infectedcells,proteinsannotatedtolocalizetoseveralorganellesofthesecretorypathwaywereoverrepresentedamongtheproteinsthatweredepl
5、etedfromthesefractionsuponinfection.Wehypothesizedthatproteins,ofwhichtheabundanceordistributionisaffectedbyinfection,arelikelytobeinvolvedintheviruslifecycle.Indeed,depletionofasmallsubsetoftheaffectedproteinsbyusingsmallinterferingRNAsidentifiedseveralho
6、stfactorsinvolvedincoronavirusinfection.TransfectionofsmallinterferingRNAstargetingeitherC11orf59orGolgiapparatusglycoprotein1resultedinincreasedvirusreplication,whereasdepletionofvesicle-traffickingproteinvesicle-traffickingproteinsec22benhancedthereleaseo
7、finfectiousprogenyvirus.Over-expressionoftheseproteins,ontheotherhand,hadanegativeeffectonvirusreplication.Overall,ourstudyshowsthattheSILACapproachisasuitabletooltostudyhost–pathogeninteractionsandtoidentifyhostproteinsinvolvedinvirusreplication.Subcellu
8、larlocalizationofidentifiedproteins;thedistributionoftheaffectedproteinswascomparedwiththatofalltheidentifiedproteins(top)andtheaffectedproteinswerealsocategorizedfortheenrichedanddepletedproteinsseparately(bottom).
