eIF2α Kinases Regulate Development through the BzpR Transcription Factor in Dictyostelium discoideum 英文参考文献.docVIP

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eIF2α Kinases Regulate Development through the BzpR Transcription Factor in Dictyostelium discoideum 英文参考文献.doc

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eIF2α Kinases Regulate Development through the BzpR Transcription Factor in Dictyostelium discoideum 英文参考文献

eIF2aKinasesRegulateDevelopmentthroughtheBzpR TranscriptionFactorinDictyosteliumdiscoideum CharlesK.Singleton*,YanhuaXiong,JanetH.Kirsten,KelseyP.Pendleton DepartmentofBiologicalSciences,VanderbiltUniversity,Nashville,Tennessee,UnitedStatesofAmerica Abstract Background: A major mechanism of translational regulation in response to a variety of stresses is mediated by phosphorylationofeIF2atoreducedeliveryofinitiatortRNAstoscanningribosomes.ForsomemRNAs,oftenencodinga bZIPtranscriptionfactor,eIF2aphosphorylationleadstoenhancedtranslationduetodelayedreinitiationatupstreamopen reading frames. Dictyostelium cells possess at least three eIF2a kinases that regulate various portions of the starvation- induceddevelopmentalprogram.CellspossessinganeIF2athatcannotbephosphorylated(BS167)showabnormalitiesin growthanddevelopment.WesoughttoidentifyabZIPproteininDictyosteliumwhoseproductioniscontrolledbytheeIF2a regulatorysystem. Principal Findings: Cells disrupted in the bzpR gene had similar developmental defects as BS167 cells, including small entities,stalkdefects,andreducedsporeviability.b-galactosidaseproductionwasusedtoexaminetranslationfrommRNA containingthebzpR59UTR.Whileproteinproductionwasreadilyapparentandregulatedtemporallyandspatiallyinwild typecells,essentiallynob-galactosidasewasproducedindevelopingBS167cellseventhoughthelacZmRNAlevelswere thesameasthoseinwildtypecells.Also,noproteinproductionwasobservedinstrainslackingIfkAorIfkBeIF2akinases. GFPfusions,withappropriateinternalcontrols,wereusedtodirectlydemonstratethatthebzpR59UTR,possessing7uORFs, suppressed translation by 12 fold. Suppression occurred even when all but one uORF was deleted, and translational suppressionwasremovedwhentheATGofthesingleuORFwasmutated. Conclusions:ThefindingsindicatethatBzpRregulatesaspectsofthedevelopmentprograminDictyostelium,servingasa downstream effector of eIF2a phosphorylation. Its production is temporally and spatially regulated by eIF2a phosphorylationbyIfkAandIfkBandthrou