Functional Characterizations of RIG-I to GCRV and ViralBacterial PAMPs in Grass Carp Ctenopharyngodon idella 英文参考文献.docVIP

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Functional Characterizations of RIG-I to GCRV and ViralBacterial PAMPs in Grass Carp Ctenopharyngodon idella 英文参考文献.doc

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Functional Characterizations of RIG-I to GCRV and ViralBacterial PAMPs in Grass Carp Ctenopharyngodon idella 英文参考文献

FunctionalCharacterizationsofRIG-ItoGCRVandViral/ BacterialPAMPsinGrassCarpCtenopharyngodonidella LijunChen,JianguoSu*,ChunrongYang,LiminPeng,QuanyuanWan,LanWang CollegeofAnimalScienceandTechnology,NorthwestAFUniversity,ShaanxiKeyLaboratoryofMolecularBiologyforAgriculture,Yangling,People’sRepublicofChina Abstract Background: RIG-I (retinoic acid inducible gene-I) is one of the key cytosolic pattern recognition receptors (PRRs) for detectingnucleotidepathogenassociatedmolecularpatterns(PAMPs)andmediatingtheinductionoftypeIinterferonand inflammatorycytokinesininnateimmuneresponse.Thoughthemechanismiswellcharacterizedinmammals,thestudyof theaccuratefunctionofRIG-Iinteleostsisstillinitsinfancy. Methodology/PrincipalFindings:ToclarifythefunctionalcharacterizationsofRIG-IingrasscarpCtenopharyngodonidella (CiRIG-I),sixrepresentativeoverexpressionplasmidswereconstructedandtransfectedintoC.idellakidney(CIK)celllinesto obtainstablyexpressingrecombinantproteins,respectively.Avirustitertestand96-wellplatestainingassayshowedthat all constructs exhibited the antiviral activity somewhat. The quantitative real-time RT-PCR (qRT-PCR) demonstrated that mRNAexpressionsofCiIPS-1,CiIFN-IandCiMx2wereregulatedbynotonlyvirus(GCRV)orviralPAMP(poly(IC))challenge butalsobacterialPAMPs(LPSandPGN)stimulationinthesteadilytransfectedcells.Theresultsshowedthatthefull-length CiRIG-IplayedakeyroleinRLRpathway.Therepressordomain(RD)exertedaninhibitoryfunctionofthesignalingchannel under all utilized challenges. Caspase activation and recruitment domains (CARDs) showed a positive role in GCRV and poly(I:C) challenge. Helicase motifs were crucial for the signaling pathway upon LPS and PGN stimulation. Interestingly, DCARDs(CARDsdeleted)showedpostivemodulationinRIG-Isignaltransduction. Conclusions/Significance:TheresultsprovidedsomenovelinsightsintoRIG-Isensingwithastrikinglybroadregulationin teleosts,respondingnotonlytothedsRNAvirusorsyntheticdsRNAbutalsobacterialPAMPs. Citation: Chen L, Su J, Yang C,