Kinetic analysis of pseudouridine formation in tRNA by archaeal Cbf5 and Pus10

dc.contributor.authorKamalampeta, Rajashekhar
dc.contributor.supervisorWieden-Kothe, Ute
dc.contributor.supervisorRoussel, Marc R.
dc.date.accessioned2014-07-17T18:21:48Z
dc.date.available2014-07-17T18:21:48Z
dc.date.issued2013
dc.degree.levelPh.Den_US
dc.degree.levelPhD
dc.descriptionxiii, 119 leaves : ill. (some col.) ; 29 cmen_US
dc.description.abstractPseudouridines (Ψ), the most common modifications in RNA, are formed by stand-alone Ψ synthases in all organisms. In addition, archaea and eukaryotes use H/ACA small ribonucleoproteins for pseudouridylation. Cbf5, the catalytic component of these complexes, can also introduce Ψ55 in archaeal tRNAs in a guide RNA-independent manner. Here, kinetic and thermodynamic analyses revealed that both Pyrococcus furiosus Nop10 and Gar1 proteins enhance the catalytic ability of Cbf5 and increase its affinity for tRNA. Pus10, representing a novel Ψ synthase family, is the in vivo archaeal tRNA Ψ55 synthase. Characterization of several Pus10 variants demonstrated the importance of the thumb loop for catalysis, a potential role of the THUMP domain in tRNA binding and a new catalytic arginine which may flip the target uridine into Pus10’s active site. The quantitative characterization of the archaeal pseudouridine synthases Cbf5 and Pus10 reported here sheds light on their cellular roles in RNA modification.en_US
dc.identifier.urihttps://hdl.handle.net/10133/3464
dc.language.isoen_CAen_US
dc.proquestyesNoen_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.subjectPseudouridine -- Researchen_US
dc.subjectTransfer RNA -- Researchen_US
dc.subjectArchaebacteria -- Researchen_US
dc.subjectDissertations, Academicen_US
dc.titleKinetic analysis of pseudouridine formation in tRNA by archaeal Cbf5 and Pus10en_US
dc.typeThesisen_US
Files
Original bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
KAMALAMPETA_RAJASHEKAR_PHD_2013.pdf
Size:
1.81 MB
Format:
Adobe Portable Document Format
Description:
License bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
license.txt
Size:
1.63 KB
Format:
Item-specific license agreed upon to submission
Description: