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dc.contributorNot Applicableen_US
dc.contributor.authorREAD, LAURIE K. Principal Investigatoren_US
dc.date30-Nov-11en_US
dc.date2010en_US
dc.date.accessioned2011-04-18T20:58:19Zen_US
dc.date.accessioned2011-04-19T18:31:01Z
dc.date.available1-Dec-05en_US
dc.date.available2011-04-18T20:58:19Zen_US
dc.date.available2011-04-19T18:31:01Z
dc.date.issued2011-04-18T20:58:19Zen_US
dc.identifier7727926en_US
dc.identifier5R01AI061580-05en_US
dc.identifier61580en_US
dc.identifier.urihttp://hdl.handle.net/10477/1037
dc.descriptionAddress;Affect;Affinity;Apocytochrome b;Area;Base Pairing;Biochemical;Biological Assay;Biological Models;Blood Circulation;Bypass;Cells;Cleaved cell;Complement;Complex;Cues;Data;Detection;Diphosphates;Down-Regulation;Enzymes;Essential Genes;Event;fascinate;Gene Expression;Genes;Genetic;genetic analysis;Goals;Guide RNA;Immunoprecipitation;In Vitro;in vitro Assay;in vivo;insertion/deletion mutation;insight;knock-down;Laboratories;Life Cycle Stages;Ligation;Mass Spectrum Analysis;Mediating;Messenger RNA;Mitochondria;Mitochondrial RNA;Modeling;mutant;Northern Blotting;novel;Physiological;Process;Production;protein expression;Proteins;Reaction;Reading;Recombinant Proteins;Recombinants;Regulation;Research Personnel;research study;response;RNA;RNA Binding;RNA Editing;RNA Interference;RNA Recognition Motif;RNA-Binding Proteins;Role;Site;Structural Protein;Structure;System;Testing;Titrations;Trypanosoma;Trypanosoma brucei brucei;Uridine;yeast two hybrid system;Yeasts;en_US
dc.descriptionAmount: $ 291373en_US
dc.description.abstractRNA editing is a novel and essential gene regulatory mechanism in trypanosomes that altersmitochondria! RNAs by specific uridine insertion and deletion to form functional mRNAs. The geneticinformation for editing is transferred from frans-acting gRNAs to mRNAs through base-pairinginteractions. While considerable progress has been made in understanding the structure andcomposition of the core editing machinery, regulatory factors that affect the accuracy and efficiencyof the editing process remain elusive. Our laboratory recently provided genetic evidence for the firstRNA editing regulatory factori'the RNA binding protein, RBP16. RNAi-mediated knock-down ofRBP16 in procyclic form (PF)trypanosomes leads to a dramatic and specific decrease in editing ofapocytochrome b (CYb) RNA. CYb gRNA abundance is unaffected in RBP16 knock-downs,suggesting that regulation takes place at the level of gRNA utilization. Consistent with its role in vivo,RBP16 reproducibly stimulates RNA editing in vitro up to 5-fold. Preliminary data suggest that RBP16 enhances editing by an intriguing two-step mechanism, one aspect of which is independent ofspecific, high affinity RBP16-RNAinteractions. Our goal is to determine the scope and mechanism ofRBP16 RNA editing regulation. In Aim 1, we will down-regulate RBP16 expression in bloodstreamform (BF)trypanosomes and analyze the effects on BF-specific and constitutive editing events. Wewill test the hypothesis that RBP16 facilitates recruitment of specific gRNAs to editosomes byanalysis of editosome-associated gRNAs in PF and BF RBP16 knock-down cells. In Aim 2, we willdirectly test the model that RBP16 stimulates both pre- and post-mRNA cleavage steps of editing.The role the RNP1 and RGG RNA binding domains of RBP16 in this process will be examined bycomparison of mutant and wild type RBP16 in the in vitro assay. In Aim 3, RBP16-interacting proteins will be identified by both expression of TAP-tagged RBP16 in PF and BF T. brucei and by ayeast two-hybrid screen. This combined genetic and biochemical approach will provide important insight into the largely unexplored area of RNA editing regulation.en_US
dc.titleREGULATION OF RNA EDITING IN TRYPANSOMA BRUCEIen_US
dc.typeNIH Grant Awarden_US


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