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Reconstitution of CFTR Ubiquitination and Endoplasmic Reticulum Quality Control

Estabrooks, Samuel K (2020) Reconstitution of CFTR Ubiquitination and Endoplasmic Reticulum Quality Control. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

Upon its identification in 1989, mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) were conclusively linked to cystic fibrosis. Many of the resultant mutant proteins were shown to misfold during synthesis and be selected for degradation by protein quality control machinery. However, small molecule “corrector” compounds help shield some variants from recognition by degradative machinery, improving protein trafficking in cell culture. Although some of these correctors have received FDA approval, their use in patients has had limited and variable efficacy. Based on emerging data, I propose that inhibition of certain E3 ubiquitin ligases— the enzymes ultimately responsible for committing CFTR to degradation— could synergize with corrector treatment to favor enhanced CFTR maturation and function. Specifically, I propose that the ubiquitin ligase CHIP is among the most attractive candidates for chemical inhibition because it is so broadly involved in the triage of misfolded CFTR substrates, contributing to the turnover of CFTR both at the endoplasmic reticulum as it is synthesized and at the cell surface where it functions. As a first step toward this goal, I have developed assays to measure ubiquitin conjugation and CHIP activity in a variety of conditions, and most notably in the presence of misfolded CFTR variants. In this thesis, I present my findings from these assays and discuss how these results further our understanding of the degradation of misfolded, disease-causing CFTR variants.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Estabrooks, Samuel Kske12@pitt.eduske12
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairBrodsky, Jeffrey Ljbrodsky@pitt.edujbrodsky
Committee MemberBomberger, Jennifer Mjbomb@pitt.edujbomb
Committee MemberBoyle, Jon Pboylej@pitt.eduboylej
Committee MemberChapman, Deborah Ldlc7@pitt.edudlc7
Committee MemberPipas, James Ppipas@pitt.edupipas
Date: 8 June 2020
Date Type: Publication
Defense Date: 8 April 2020
Approval Date: 8 June 2020
Submission Date: 9 April 2020
Access Restriction: 1 year -- Restrict access to University of Pittsburgh for a period of 1 year.
Number of Pages: 174
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: protein quality control; cystic fibrosis; CFTR; ERAD; ubiquitination; ubiquitin ligases; CHIP
Date Deposited: 08 Jun 2020 16:11
Last Modified: 08 Jun 2020 16:11
URI: http://d-scholarship.pitt.edu/id/eprint/38655

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