Role of Bunyamwera Orthobunyavirus NSs Protein in Infection of Mosquito Cells 英文参考文献.docVIP

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Role of Bunyamwera Orthobunyavirus NSs Protein in Infection of Mosquito Cells 英文参考文献.doc

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Role of Bunyamwera Orthobunyavirus NSs Protein in Infection of Mosquito Cells 英文参考文献

RoleofBunyamweraOrthobunyavirusNSsProteinin InfectionofMosquitoCells AgnieszkaM.Szemiel1,Anna-BellaFailloux2,RichardM.Elliott1* 1BiomedicalSciencesResearchComplex,SchoolofBiology,UniversityofSt.Andrews,NorthHaugh,St.Andrews,Scotland,UnitedKingdom,2DepartmentofVirology, InstitutPasteur,Paris,France Abstract Background:BunyamweraorthobunyavirusisboththeprototypeandstudymodeloftheBunyaviridaefamily.TheviralNSs proteinseemstocontributetothedifferentoutcomesofinfectioninmammalianandmosquitocelllines.However,only limited information is available on the growth of Bunyamwera virus in cultured mosquito cells other than the Aedes albopictusC6/36line. MethodologyandPrincipalFindings:TodeterminepotentialfunctionsoftheNSsproteininmosquitocells,replicationof wild-typevirusandarecombinantNSsdeletionmutantwascomparedinAe.albopictusC6/36,C7-10andU4.4cells,andin Ae. aegypti Ae cells by monitoring N protein production and virus yields at various times post infection. Both viruses establishedpersistentinfections,withtheexceptionofNSsdeletionmutantinU4.4cells.TheNSsproteinwasnonessential for growth in C6/36 and C7-10 cells, but was important for productive replication in U4.4 and Ae cells. Fluorescence microscopystudiesusingrecombinantvirusesexpressinggreenfluorescentproteinallowedobservationofthreestagesof infection,early,acuteandlate,duringwhichinfectedcellsunderwentmorphologicalchanges.IntheabsenceofNSs,these changeswerelesspronounced.AnRNAiresponseefficientlyreducedvirusreplicationinU4.4cellstransfectedwithvirus specificdsRNA,butnotinC6/36orC7/10cells.Lastly,Ae.aegyptimosquitoeswereexposedtoblood-mealcontainingeither wild-type or NSs deletion virus, and at various times post-feeding, infection and disseminated infection rates were measured. Compared to wild-type virus, infection rates by the mutant virus were lower and more variable. If the NSs deletionviruswasabletoestablishinfection,itwasdetectedinsalivaryglandsat6dayspost-infection,3dayslaterthan wild-typevirus. Conclusions/Si