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dc.contributor.advisorKingston, Robert
dc.contributor.advisorWalker, Suzanne
dc.contributor.authorLevine, Zebulon Gehrett
dc.date.accessioned2019-12-12T09:27:59Z
dash.embargo.terms2021-05-01
dc.date.created2019-05
dc.date.issued2019-05-15
dc.date.submitted2019
dc.identifier.citationLevine, Zebulon Gehrett. 2019. Strategies to dissect O-GlcNAc Transferase biochemistry in vitro and in cells. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences.
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:42029809*
dc.description.abstractIn this dissertation, I develop tools to allow us to understand how O-GlcNAc Transferase (OGT) recognizes its substrates and to separate OGT's catalytic functions in a cellular context. In the introduction, I introduce OGT's multiple catalytic activities and examine why they are necessary for mammalian cell survival. In the second chapter, I use targeted and high-throughput methods to determine the role of OGT's N-terminal tetratricopeptide repeat (TPR) domain in substrate selection. I demonstrate that the majority of substrate are bound in the lumen of this superhelical domain, with conserved Asn binding the peptide backbone and key Asp residues playing a selectivity-determining role. In the third chapter, I develop a system for replacing OGT with enzyme variants in living cells; this system allows me to conclude that O-GlcNAc transfer to Ser/Thr, not HCF-1 cleavage, is essential for cell viability. Together, these chapters present methods that can be applied for both further biochemical studies of OGT substrate selection as well as future OGT structure-function studies in cells. Last, I present an appendix on by-residue conservation of OGT that may guide such future studies.
dc.description.sponsorshipChemical Biology
dc.format.mimetypeapplication/pdf
dc.language.isoen
dash.licenseLAA
dc.subjectO-GlcNAc, enzymology, biochemistry, rapid degradation, substrate recognition, OGT
dc.titleStrategies to dissect O-GlcNAc Transferase biochemistry in vitro and in cells
dc.typeThesis or Dissertation
dash.depositing.authorLevine, Zebulon Gehrett
dash.embargo.until2021-05-01
dc.date.available2019-12-12T09:27:59Z
thesis.degree.date2019
thesis.degree.grantorGraduate School of Arts & Sciences
thesis.degree.grantorGraduate School of Arts & Sciences
thesis.degree.levelDoctoral
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy
thesis.degree.nameDoctor of Philosophy
dc.contributor.committeeMemberMitchison, Timothy
dc.contributor.committeeMemberKiessling, Laura
dc.type.materialtext
thesis.degree.departmentChemical Biology
thesis.degree.departmentChemical Biology
dash.identifier.vireo
dc.identifier.orcid0000-0002-0830-9032
dash.author.emailzebulon.levine@gmail.com


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