Detection of pseudouridine synthase activity with a fluorescence-based yeast reporter system

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dc.contributor.authorDueck, Elijah
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorKothe, Ute
dc.date.accessioned2021-01-15T16:10:28Z
dc.date.available2021-01-15T16:10:28Z
dc.date.issued2020
dc.degree.levelMastersen_US
dc.description.abstractPseudouridine is an isomer of uridine and a post-transcriptional RNA modification in all domains of life. Its formation is catalyzed by pseudouridine synthases functioning as standalone proteins or protein-RNA complexes called H/ACA small nucleolar ribonucleoproteins (H/ACA snoRNPs). Whereas pseudouridylation was long known to assist in stabilizing noncoding RNAs, it was discovered also in mRNAs in 2014. mRNA pseudouridylation varies in response to stress, and artificial pseudouridylation of stop codons facilitates nonsense suppression. However, the functional significance of mRNA pseudouridylation and the regulation of the pseudouridine synthases responsible are only partially understood. To study mRNA pseudouridylation by specific pseudouridine synthases, I developed a fluorescence-based reporter system in S. cerevisiae and used it to assay for standalone Pus7, snR5 and snR81 H/ACA snoRNP activity. This novel system detects snR5 H/ACA snoRNP activity and can be used to further investigate the role of pseudouridine synthases in gene expression.en_US
dc.identifier.urihttps://hdl.handle.net/10133/5823
dc.language.isoen_USen_US
dc.proquest.subjectBiochemistry [0487]en_US
dc.proquest.subjectMolecular biology [0307]en_US
dc.proquest.subjectCellular biology [0379]en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta. : University of Lethbridge, Dept. of Chemistry & 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.subjectDissertations, Academicen_US
dc.subjectFluorescence microscopyen_US
dc.subjectGene expressionen_US
dc.subjectMessenger RNAen_US
dc.subjectPseudouridineen_US
dc.subjectSaccharomyces cerevisiaeen_US
dc.titleDetection of pseudouridine synthase activity with a fluorescence-based yeast reporter systemen_US
dc.typeThesisen_US
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