Functions of the Nonsense-Mediated mRNA Decay Pathway in Drosophila Development 英文参考文献.docVIP

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Functions of the Nonsense-Mediated mRNA Decay Pathway in Drosophila Development 英文参考文献.doc

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Functions of the Nonsense-Mediated mRNA Decay Pathway in Drosophila Development 英文参考文献

FunctionsoftheNonsense-MediatedmRNA DecayPathwayinDrosophilaDevelopment Mark M.Metzstein¤,Mark A.Krasnow* HowardHughesMedicalInstituteandDepartmentofBiochemistry,StanfordUniversitySchoolofMedicine,Stanford,California,UnitedStatesofAmerica Nonsense-mediated mRNA decay (NMD) is a cellular surveillance mechanism that degrades transcripts containing prematuretranslationterminationcodons,anditalsoinfluencesexpressionofcertainwild-typetranscripts.Although thebiochemicalmechanismsofNMDhavebeenstudiedintensively,itsdevelopmentalfunctionsandimportanceare lessclear.Here,wedescribetheisolationandcharacterizationofDrosophila‘‘photoshop’’mutations,whichincrease expressionofgreenfluorescentproteinandothertransgenes.Mappingandmolecularanalysesshowthatphotoshop mutationsareloss-of-functionmutationsintheDrosophilahomologsofNMDgenesUpf1,Upf2,andSmg1.Wefindthat Upf1andUpf2arebroadlyactiveduringdevelopment,andtheyarerequiredforNMDaswellasforproperexpression ofdozensofwild-typegenesduringdevelopmentandforlarvalviability.GeneticmosaicanalysisshowsthatUpf1and Upf2arerequiredforgrowthand/orsurvivalofimaginalcellclones,butthisdefectcanbeovercomeifsurrounding wild-typecellsareeliminated.Bycontrast,wefindthatthePI3K-relatedkinaseSmg1potentiatesbutisnotrequired for NMD or for viability, implying that the Upf1 phosphorylation cycle that is required for mammalian and CaenorhabditiselegansNMDhasamorelimitedroleduringDrosophiladevelopment.Finally,weshowthattheSV4039 UTR,presentinmanyDrosophilatransgenes,targetsthetransgenesforregulationbytheNMDpathway.Theresults establishthattheDrosophilaNMDpathwayisbroadlyactiveandessentialfordevelopment,andonecriticalfunction ofthepathwayistoendowproliferatingimaginalcellswithacompetitivegrowthadvantagethatpreventsthemfrom beingovertakenbyotherproliferatingcells. Citation:MetzsteinMM,KrasnowMA(2006)Functionsofthenonsense-mediatedmRNAdecaypathwayinDrosophiladevelopment.PLoSGenet2(12):e180.doi:10.1371/ journal.pgen.0020180 ylation cycle (reviewed in [13]). Upf1