Alternative Splicing Regulation During C. elegans Development Splicing Factors as Regulated Targets 英文参考文献.docVIP

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Alternative Splicing Regulation During C. elegans Development Splicing Factors as Regulated Targets 英文参考文献.doc

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Alternative Splicing Regulation During C. elegans Development Splicing Factors as Regulated Targets 英文参考文献

AlternativeSplicingRegulationDuringC.elegans Development:SplicingFactorsasRegulatedTargets SergioBarberan-Soler1,2,AlanM.Zahler1,2 * 1DepartmentofMCDBiology,UniversityofCaliforniaSantaCruz,SantaCruz,California,UnitedStatesofAmerica,2CenterforMolecularBiologyofRNA,Universityof CaliforniaSantaCruz,SantaCruz,California,UnitedStatesofAmerica Abstract Alternativesplicinggeneratesproteindiversityandallowsforpost-transcriptionalgeneregulation.Estimatessuggestthat 10%ofthegenesinCaenorhabditiselegansundergoalternativesplicing.Weconstructedasplicing-sensitivemicroarrayto detect alternative splicing for 352 cassette exons and tested for changes in alternative splicing of these genes during development.Wefoundthatthemicroarraydatapredictedthat62/352(,18%)ofthealternativesplicingeventsstudied show a strong change in the relative levels of the spliced isoforms (.4-fold) during development. Confirmation of the microarray data by RT-PCR was obtained for 70% of randomly selected genes tested. Among the genes with the most developmentallyregulatedalternativelysplicingwasthehnRNPF/Hsplicingfactorhomolog,W02D3.11–nownamedhrpf- 1.Forthecassetteexonofhrpf-1,theinclusionisoformcomprises65%ofhrpf-1steadystatemessagesinembryosbutonly 0.1% in the first larval stage. This dramatic change in the alternative splicing of an alternative splicing factor suggests a complexcascadeofsplicingregulationduringdevelopment.Weanalyzedsplicinginembryosfromastrainwithamutation in the splicing factor sym-2, another hnRNP F/H homolog. We found that approximately half of the genes with large alternativesplicingchangesbetweentheembryoandL1stagesareregulatedbysym-2inembryos.Ananalysisoftherole ofnonsense-mediateddecayinregulatingsteady-statealternativemRNAisoformswasperformed.Wefoundthat8%ofthe 352eventsstudiedhavealternativeisoformswhoserelativesteady-statelevelsinembryoschangemorethan4-foldina nonsense-mediateddecaymutant,includinghrpf-1.Strikingly,53%ofthesealternativesplicingeventsthatareaffectedby NMDinou