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- W2728549966 abstract "IntheprotozoanparasiteTrypanosomabruceigeneexpressionismostlyregulatedonthepost-‐transcriptionallevel.C3Hzincfingershaveshowntoplayanimportantroleinthisprocess.Sofar,alldescribedC3HzincfingerinTrypanosomabruceihaveshowntostabilizetheirtargets.ThisworkfocusesonthezincfingerproteinZC3H32.Alarge-‐scaleisolationoffreeandmembrane-‐boundpolyribosomesfromproyclicandbloodstreamtrypanosomes,whichisalsodescribedinthisthesis,suggestedthatthisproteinassociateswithpolyribosomes.However,small-‐scalepolyribosomeisolationsfollowedbyWesternblottingrevealedthatonlyaminorfractionofitisactuallyassociatedwithpolyribosomes.ImmunofluorescenceshowedthatZC3H32localizestothecytoplasm.Ayeast-‐two-‐hybridscreenidentifieditasaputativebindingpartnerofMKT1.Thisinteractionwasconfirmedbyco-‐immunoprecipitation.RNAi,aswellasknock-‐outstudies,showedthatZC3H32isessentialinthebloodstreamform.Northernblotting,aswellasaSILACscreenbyUrbaniaketal.indicatethatitisalsoenrichedinthislife-‐cyclestageascomparedtoprocyclics.ArtificialtetheringofZC3H32toareporterRNAleadtotheRNA’sdegradation,suggestingthatthisproteinhasadestabilizingeffect.TetheringofZC3H32fragmentsrevealedthatbothitsN-‐terminal,aswellasitsC-‐terminalregionareabletogeneratethisdestabilizationeffect,whilethemiddleregion,containingthezincfingerdomains,can’t.RNAisolationfrompolyribosomalfractionsshowedthattetheringofZC3H32alsodecreasesthetranslationofthereporterRNA.High-‐throughputsequencingofpoly-‐A+RNAfromaZC3H32-‐RNAicelllinerevealed20RNAsthatwereup-‐regulateduponZC3H32knock-‐downandthusmightbeputativetargets.TheupregulationwasconfirmedforthreeofthesecandidateRNAs.ThemajorityoftheputativeZC3H32targetsplayaroleinthetrypanosome’senergymetabolismand15ofthemareup-‐regulatedinprocyclics.TheseresultssuggestthatZC3H32mightbeinvolvedinthestage-‐specificregulationoftheseRNAsinthebloodstreamform." @default.
- W2728549966 created "2017-07-14" @default.
- W2728549966 creator A5018068804 @default.
- W2728549966 date "2014-01-01" @default.
- W2728549966 modified "2023-09-23" @default.
- W2728549966 title "The role of ZC3H32 in Trypanosoma brucei" @default.
- W2728549966 doi "https://doi.org/10.11588/heidok.00017057" @default.
- W2728549966 hasPublicationYear "2014" @default.
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