Functional and mechanistic characterization of two tRNA modifying enzymes

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dc.contributor.authorKeffer-Wilkes, Laura C.
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorWieden-Kothe, Ute
dc.date.accessioned2017-03-15T20:21:33Z
dc.date.available2017-03-15T20:21:33Z
dc.date.issued2016
dc.degree.levelPh.Den_US
dc.description.abstractThe formation of pseudouridine and 5-methyluridine (m5U) in the T-arm of transfer RNAs (tRNAs) is near-universally conserved. These two modifications are formed in Escherichia coli by the pseudouridine synthase TruB and the S-adenosylmethionine-dependent methyltransferase TrmA, respectively. In this thesis, I investigate the function and mechanisms of these two tRNA modifying enzymes. First, in vitro and in vivo analysis of TruB reveals that this enzyme is acting as a tRNA chaperone which proves a long outstanding hypothesis. Secondly, characterization of ligand binding by TrmA shows that binding is cooperative and disruption of tRNA elbow region tertiary interactions by TrmA is essential for efficient tRNA binding and catalysis, leading to future analysis. In conclusion, my studies further our understanding of the mechanism and function of tRNA modifications and modifying enzymes, as well as shed light on why all cells invest substantial resources into fine-tuning the chemical composition of tRNAs.en_US
dc.embargoNoen_US
dc.embargo.end2017-02-01
dc.embargo.start2016-09-01
dc.identifier.urihttps://hdl.handle.net/10133/4797
dc.language.isoen_USen_US
dc.proquest.subject0487en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta : University of Lethbridge, Dept. of Chemistry and Biochemistryen_US
dc.publisher.departmentDepartment of Chemistry and Biochemistryen_US
dc.publisher.facultyArts and Scienceen_US
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)en_US
dc.subjecttRNAen_US
dc.subjectPseudouridineen_US
dc.subject5-methyluridineen_US
dc.subjectChaperoneen_US
dc.subjectModificationen_US
dc.titleFunctional and mechanistic characterization of two tRNA modifying enzymesen_US
dc.typeThesisen_US
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