000042360 001__ 42360
000042360 005__ 20240124114341.0
000042360 0247_ $$2doi$$a10.6083/bpxhc42360
000042360 037__ $$aETD
000042360 041__ $$aeng
000042360 245__ $$aStrangers in a strange land: bacteria provide mechanistic insight into eukaryotic ubiquitination
000042360 260__ $$bOregon Health and Science University
000042360 269__ $$a2023-11-24
000042360 336__ $$aDissertation
000042360 502__ $$bPh.D.
000042360 502__ $$gMolecular Microbiology & Immunology
000042360 520__ $$aHere, I present a comprehensive assay termed UbiReal for monitoring each step of the Ub signaling cascade, including linkage-specific polyUb ligation and deubiquitination. Next, I use bioinformatic tools and recombinant protein expression to identify additional, HECT-like ligases from human and plant pathogens, and validate these ligases using UbiReal. I apply structural biology and biochemical techniques to elucidate the mechanism of Ub ligation by the bacterial HECT-like ligases, and capture a rare glimpse of a linkage-specific polyUb ligation interface. I leverage insights from the linkage-specific polyUb interface to engineer an NleL specific for K6-linked polyUb, and show that similar rules influence the ligation of K6-linked polyUb in humans. Finally, through analysis of a family of deubiquitinating enzymes in bacteria, I demonstrate the utility of the deubiquitinase LotA as a tool for detecting the presence of K6-linked polyUb. Collectively, this dissertation provides the critical tools for understanding K6-linked polyUb biology, and in particular sets the stage for elucidating the enigmatic role of K6-linked polyUb at the host-pathogen interface.
000042360 536__ $$oNIGMS$$cR35GM142486
000042360 540__ $$fCC BY
000042360 542__ $$fIn copyright - single owner
000042360 650__ $$aBiochemistry$$015549
000042360 650__ $$aMicrobiology$$022259
000042360 650__ $$aCrystallography, X-Ray$$030746
000042360 6531_ $$aubiquitin
000042360 6531_ $$ae3 ubiquitin ligase
000042360 6531_ $$adeubiquitinase
000042360 6531_ $$abacteria
000042360 6531_ $$anlel
000042360 6531_ $$asopa
000042360 6531_ $$alota
000042360 691__ $$aSchool of Medicine$$041369
000042360 692__ $$aDepartment of Molecular Microbiology and Immunology$$041429
000042360 7001_ $$aFranklin, Tyler$$uOregon Health and Science University$$041354$$10000-0002-0311-1190
000042360 8564_ $$yPermission document$$9606e59b7-433c-470a-a16a-5d152dcc941f$$s129731$$uhttps://digitalcollections.ohsu.edu/record/42360/files/permission-LotA.pdf
000042360 8564_ $$yPermission document$$96546c701-2746-4f2e-b9b9-64e487c5b933$$s129634$$uhttps://digitalcollections.ohsu.edu/record/42360/files/permission-PPUbireal.pdf
000042360 8564_ $$918c1389b-fcfc-492c-bc34-fcb2fd3efd30$$s6825093$$uhttps://digitalcollections.ohsu.edu/record/42360/files/Franklin.Tyler.2023.pdf
000042360 909CO $$ooai:digitalcollections.ohsu.edu:42360$$pstudent-work
000042360 980__ $$aTheses and Dissertations
000042360 981__ $$aPublished$$b2023-11-27