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dc.contributor.advisorYao, Tingting
dc.contributor.authorHazlett, Zachary S.
dc.contributor.committeememberCohen, Robert
dc.contributor.committeememberPeersen, Olve
dc.contributor.committeememberDiPietro, Santiago
dc.contributor.committeememberKennan, Alan
dc.date.accessioned2021-01-11T11:20:24Z
dc.date.available2022-01-08T11:20:24Z
dc.date.issued2020
dc.description2020 Fall.
dc.descriptionIncludes bibliographical references.
dc.description.abstractThe ubiquitin (Ub) C-terminal hydrolase, Uch37, can be found associated with the 26S proteasome as well as the INO80 chromatin remodeling complex. Bound to the 26S proteasome, it assists in regulating the degradation of Ub modified proteins. The proteasomal subunit Rpn13 binds Uch37, anchors it to the proteasome 19S regulatory particle and enhances the deubiquitinating enzyme's (DUB's) catalytic activity. While the structure of the Uch37/Rpn13 complex bound to a single Ub molecule has been characterized, much still remains unknown regarding the enzyme's substrate specificity, the molecular basis for its substrate specificity, and its function in the regulation of proteasomal degradation. In this thesis we characterize the substrate specificity of Uch37 with and without its proteasomal binding partner Rpn13. By synthesizing poly-Ub chains of various linkage types and topologies and using these Ub chains in in vitro deubiquitination assays, we were able to determine that Uch37/Rpn13 selectively cleaves branched Ub chains. This provides evidence to suggest that Uch37 is the first enzyme with activity specific for branched Ub chains. Branched Ub chains have been identified endogenously and have roles connected to the regulation of nascent misfolded polypeptides, cell cycle control, and the enhancement of proteasomal degradation. The work presented here sets out to characterize the molecular mechanism of branched chain hydrolysis by Uch37 and its binding partner Rpn13, determine the kinetics of this enzymatic reaction, and establish a system for probing the function of "debranching" by Uch37 in proteasomal degradation. The conclusion of our work builds our understanding of the complex system of intracellular signaling by Ub and unveils key elements to the primary system responsible for regulating cellular protein homeostasis.
dc.format.mediumborn digital
dc.format.mediummasters theses
dc.identifierHazlett_colostate_0053N_16389.pdf
dc.identifier.urihttps://hdl.handle.net/10217/219567
dc.languageEnglish
dc.publisherColorado State University. Libraries
dc.relation.ispartof2020- CSU Theses and Dissertations
dc.rightsCopyright of the original work is retained by the author.
dc.rights.accessEmbargo Expires: 01/08/2022
dc.subjectproteasome
dc.subjectbranched ubiquitin
dc.subjectUch37
dc.titleCharacterization of the selective hydrolysis of branched ubiquitin chains by Uch37 and its activator Rpn13
dc.typeText
dcterms.embargo.expires2022-01-08
dcterms.rights.dplaThe copyright and related rights status of this Item has not been evaluated (https://rightsstatements.org/vocab/CNE/1.0/). Please refer to the organization that has made the Item available for more information.
thesis.degree.disciplineBiochemistry and Molecular Biology
thesis.degree.grantorColorado State University
thesis.degree.levelMasters
thesis.degree.nameMaster of Science (M.S.)


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