dc.creator | Cselenyi, Christopher Stephen | |
dc.date.accessioned | 2020-08-22T17:43:40Z | |
dc.date.available | 2010-07-26 | |
dc.date.issued | 2008-07-26 | |
dc.identifier.uri | https://etd.library.vanderbilt.edu/etd-07212008-143939 | |
dc.identifier.uri | http://hdl.handle.net/1803/13363 | |
dc.description.abstract | Wnt/beta-catenin signaling controls various cell fates in metazoan development and is misregulated in several cancers and developmental disorders. Binding of a Wnt ligand to its transmembrane coreceptors, Frizzled (Fz) and LRP5/6, inhibits phosphorylation and degradation of the transcriptional coactivator beta-catenin, which then translocates to the nucleus to regulate target gene expression. To understand how Wnt signaling prevents beta-catenin degradation, I focused on the Wnt coreceptor LRP6, which is required for signal transduction and is sufficient to activate Wnt signaling when overexpressed. LRP6 has been proposed to stabilize beta-catenin by stimulating degradation of Axin, a scaffold protein required for beta-catenin degradation. In certain systems, however, Wnt-mediated Axin turnover is not detected until after beta-catenin has been stabilized. Thus, LRP6 may also signal through a mechanism distinct from Axin degradation. To establish a biochemically tractable system to test this hypothesis, I expressed and purified the LRP6 intracellular domain from bacteria and show that it promotes beta-catenin stabilization and Axin degradation in Xenopus egg extract. Using an Axin mutant that does not degrade in response to LRP6, I demonstrate that LRP6 can stabilize beta-catenin in the absence of Axin turnover. Through experiments in egg extract and reconstitution with purified proteins, I identify a mechanism whereby LRP6 stabilizes beta-catenin independently of Axin degradation by directly inhibiting GSK3's phosphorylation of beta-catenin.
In addition to studies of LRP6, I explore the role of the other Wnt coreceptor Fz, which has been suggested to be a G protein coupled receptor. Through biochemical studies in Xenopus egg extract, I demonstrate that Galphao, Galphai, Galphaq, and Gbetagamma promote beta-catenin stabilization by inhibiting GSK3’s phosphorylation of beta-catenin.
Independently of studies on Wnt signaling, I find that two enzymes involved in glycosylation, NAGK and DPAGT1, regulate anteroposterior patterning in Xenopus embryogenesis. I discover that these enzymes involved in N-glycosylation specifically regulate FGF-mediated events in Xenopus development. Because partial loss-of-function mutations in global regulators of N-glycosylation cause a group of human developmental disorders called Congenital Disorders of Glycosylation (CDGs), I suggest the use of Xenopus as a model organism to study the molecular etiology of CDGs. | |
dc.format.mimetype | application/pdf | |
dc.subject | LRP6 | |
dc.subject | G proteins | |
dc.subject | NAGK | |
dc.subject | development | |
dc.subject | Glycosylation | |
dc.subject | Xenopus | |
dc.subject | Axin | |
dc.subject | GSK3 | |
dc.subject | Wnt | |
dc.subject | Diseases -- Animal models | |
dc.title | Receptor-Mediated Activation of Canonical Wnt Signaling | |
dc.type | dissertation | |
dc.contributor.committeeMember | Susan Wente | |
dc.contributor.committeeMember | Jennifer Pietenpol | |
dc.contributor.committeeMember | Chin Chiang | |
dc.contributor.committeeMember | Ela Knapik | |
dc.contributor.committeeMember | Ethan Lee, Advisor | |
dc.type.material | text | |
thesis.degree.name | PHD | |
thesis.degree.level | dissertation | |
thesis.degree.discipline | Cell and Developmental Biology | |
thesis.degree.grantor | Vanderbilt University | |
local.embargo.terms | 2010-07-26 | |
local.embargo.lift | 2010-07-26 | |
dc.contributor.committeeChair | Robert Coffey | |