Advisory Committee Chair
Janet Collawn
Advisory Committee Members
Casey Morrow
Scott Parker
Laura Timares
Casey Weaver
Document Type
Dissertation
Date of Award
2007
Degree Name by School
Doctor of Philosophy (PhD) Heersink School of Medicine
Abstract
Cystic fibrosis (CF) is caused by mutation of one protein, the cystic fibrosis transmembrane conductance regulator (CFTR), which normally functions as a chloride channel at the apical surface of epithelial cells. The most common CFTR mutation results in the deletion of a single amino acid (phenylalanine) at position 508, which causes the protein to fold improperly. The ΔF508 CFTR is a temperature-sensitive (TS) processing mutant: At the restrictive temperature, 37°C, ΔF508 CFTR misfolds in the endoplasmic reticulum (ER) and is degraded, but at the permissive temperature, 27°C, it is “rescued” from degradation (rΔF508). In particular, rΔF508 CFTR folds correctly enough to exit the ER and produces a chloride channel at the cell surface, similar to the wild-type (WT) protein. Unfortunately, rΔF508 is rapidly degraded after returning to the restrictive temperature, suggesting a surface stability defect. Therefore, it is clear that the ΔF508 CFTR defect can be corrected under certain conditions, but first we must understand how the WT and ΔF508 CFTR proteins are handled differentially by the ER and at the cell surface, which is the aim of the work presented herein. We first characterized an interaction between CFTR and a protein associated with ER degradation, valosin-containing protein (VCP), and found that it interacts with ΔF508 CFTR, but not with the WT protein. This study provides further evidence that WT CFTR is processed efficiently in the ER. Second, with regard to ΔF508 CFTR, we found that culture at the permissive temperature corrects not only the folding defect in the ER, but also the impaired surface stability of ΔF508 CFTR. This result iv suggests that, like the folding defect, the surface stability defect of ΔF508 CFTR is also TS. Additionally, we showed that two small molecular compounds known to mimic permissive temperature culture of ΔF508 by correcting its ER processing defect also enhance its surface stability at 37°C. These studies add significantly to current CF knowledge and may aid in the design of future therapeutics for CF and other diseases resulting from protein folding mutations. Specifically, we identify specific differences between ER processing and surface trafficking of WT and ΔF508 CFTR, and we reveal a novel mode of action for two known pharmacological interventions.
Recommended Citation
Goldstein, Rebecca F., "Protein Interaction And Cell Surface Trafficking Differences Between Wild-Type And ΔF508 Cystic Fibrosis Transmembrane Conductance Regulator" (2007). All ETDs from UAB. 3706.
https://digitalcommons.library.uab.edu/etd-collection/3706