title: CFTR expression regulation by the unfolded protein response.
abstract: The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel and key regulator of epithelial functions. Mutations in the CFTR gene lead to reduced or dysfunctional CFTR protein and cause cystic fibrosis (CF), a generalized exocrinopathy affecting multiple organs. In the airways, loss of CFTR function leads to thickened mucus, reduced mucociliary clearance, chronic infections, and respiratory failure. Common airway disorders such as bronchitis and chronic obstructive pulmonary disease (COPD) also present CF-like symptoms such as mucus congestion and chronic inflammation without mutations in CFTR. The primary risk factors for COPD and chronic bronchitis include environmental stress insults such as pollutants and infections that often result in hypoxic conditions. Furthermore, environmental factors such as cigarette smoke and reactive oxygen species have been implicated in reduced CFTR function. Activation of cellular stress responses by these factors promotes differential, stress-associated gene expression regulation. During our investigations on the mechanisms of CFTR expression regulation, we have shown that the ER stress response, the unfolded protein response (UPR), decreases CFTR expression at the transcriptional, translational, and maturational levels. Here, we provide a detailed description of the methods we employ to study CFTR expression regulation by the UPR. Similar approaches are applicable in studies on other genes and how they are affected by the UPR.
authors.affiliation: Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Rafal
authors.initials: R
authors.lastname: Bartoszewski
authors.forename: Andras
authors.initials: A
authors.lastname: Rab
authors.forename: Lianwu
authors.initials: L
authors.lastname: Fu
authors.forename: Sylwia
authors.initials: S
authors.lastname: Bartoszewska
authors.forename: James
authors.initials: J
authors.lastname: Collawn
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Bebok
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D047369:Chromatin Immunoprecipitation / Q000379:methods; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000235:genetics* / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D005821:Genetic Techniques; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014158:Transcription, Genetic; D056811:Unfolded Protein Response
chemical_list: D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator