NOX2β A Novel Splice Variant of NOX2 That Regulates NADPH Oxidase Activity in Macrophages 英文参考文献.docVIP

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NOX2β A Novel Splice Variant of NOX2 That Regulates NADPH Oxidase Activity in Macrophages 英文参考文献.doc

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NOX2β A Novel Splice Variant of NOX2 That Regulates NADPH Oxidase Activity in Macrophages 英文参考文献

NOX2b:ANovelSpliceVariantofNOX2ThatRegulates NADPHOxidaseActivityinMacrophages CraigB.Harrison1.,StavrosSelemidis1*.,ElizabethGuida1,PaulT.King2,ChristopherG.Sobey1, GrantR.Drummond1 1VascularBiologyandImmunopharmacologyGroup,DepartmentofPharmacology,MonashUniversity,Clayton,Victoria,Australia,2DepartmentofMedicine/Respiratory Medicine,MonashMedicalCentre,Clayton,Victoria,Australia Abstract Nox2oxidaseisoneisoforminafamilyofsevenNADPHoxidasesthatgeneratereactiveoxygenspecies(ROS)andthereby contribute to physiological and pathological processes including host defense, redox signaling and oxidative tissue damage. While alternative mRNA splicing has been shown to influence the activity of several Nox-family proteins, functionallyrelevantsplicevariantsofNox2havenotpreviouslybeenidentified.Weimmunoscreenedseveralmousetissues andcellsforthepresenceoftruncatedNox2proteinsandidentifieda30kDaproteininlung,spleenandmacrophages.RT- PCR analysis of mRNA from primary and immortalised (RAW264.7) mouse macrophages, and from human alveolar macrophages,identifiedatruncatedNox2transcriptwhich,uponsequenceanalysis,wasfoundtobeaproductofthe‘exon skipping’modeofalternativesplicing,lackingexons4–10oftheNox2gene.Thepredictedproteiniscomparableinsizeto thatidentifiedbyimmunoscreeningandcontainstwotransmembranehelicesandanextendedcytosolicC-terminuswith bindingsitesforNADPHandtheNoxorganiserproteinp47phox.Importantly,selectivesiRNA-mediatedknockdownofthe transcript reduced expression of the 30kDa protein in macrophages, and suppressed phorbol ester-stimulated ROS productionby50%.WethusprovidethefirstevidencethatNox2undergoesalternativemRNAsplicingtoyielda30kDa protein – herein termed Nox2b – that regulates NADPH oxidase activity in macrophages from mice and humans. The discoveryofNox2bpavesthewayforfutureexaminationofitsroleinphysiologicalandpathologicalprocesses. Citation:HarrisonCB,SelemidisS,GuidaE,KingPT,SobeyCG,etal.(2012)NOX2b:ANovelSpliceVariantofNOX2ThatRegulatesNADPHOxidaseActivi