Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/107869
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Type: Journal article
Title: Ribosomal stalk protein silencing partially corrects the ΔF508-CFTR functional expression defect
Other Titles: Ribosomal stalk protein silencing partially corrects the DeltaF508-CFTR functional expression defect
Author: Veit, G.
Oliver, K.
Apaja, P.
Perdomo, D.
Bidaud-Meynard, A.
Lin, S.
Guo, J.
Icyuz, M.
Sorscher, E.
Hartman IV, J.
Lukacs, G.
Citation: PLoS Biology, 2016; 14(5):e1002462-1-e1002462-32
Publisher: Public Library of Science (PLoS)
Issue Date: 2016
ISSN: 1544-9173
1545-7885
Editor: Ron, D.
Statement of
Responsibility: 
Guido Veit, Kathryn Oliver, Pirjo M. Apaja, Doranda Perdomo, Aurélien Bidaud-Meynard, Sheng-Ting Lin, Jingyu Guo, Mert Icyuz, Eric J. Sorscher, John L. Hartman IV, Gergely L. Lukacs
Abstract: The most common cystic fibrosis (CF) causing mutation, deletion of phenylalanine 508 (ΔF508 or Phe508del), results in functional expression defect of the CF transmembrane conductance regulator (CFTR) at the apical plasma membrane (PM) of secretory epithelia, which is attributed to the degradation of the misfolded channel at the endoplasmic reticulum (ER). Deletion of phenylalanine 670 (ΔF670) in the yeast oligomycin resistance 1 gene (YOR1, an ABC transporter) of Saccharomyces cerevisiae phenocopies the ΔF508-CFTR folding and trafficking defects. Genome-wide phenotypic (phenomic) analysis of the Yor1-ΔF670 biogenesis identified several modifier genes of mRNA processing and translation, which conferred oligomycin resistance to yeast. Silencing of orthologues of these candidate genes enhanced the ΔF508-CFTR functional expression at the apical PM in human CF bronchial epithelia. Although knockdown of RPL12, a component of the ribosomal stalk, attenuated the translational elongation rate, it increased the folding efficiency as well as the conformational stability of the ΔF508-CFTR, manifesting in 3-fold augmented PM density and function of the mutant. Combination of RPL12 knockdown with the corrector drug, VX-809 (lumacaftor) restored the mutant function to ~50% of the wild-type channel in primary CFTRΔF508/ΔF508 human bronchial epithelia. These results and the observation that silencing of other ribosomal stalk proteins partially rescue the loss-of-function phenotype of ΔF508-CFTR suggest that the ribosomal stalk modulates the folding efficiency of the mutant and is a potential therapeutic target for correction of the ΔF508-CFTR folding defect.
Keywords: Bronchi
Cells, Cultured
Epithelial Cells
Yeasts
Cystic Fibrosis
Aminopyridines
Peptide Elongation Factor 2
ATP-Binding Cassette Transporters
Cystic Fibrosis Transmembrane Conductance Regulator
Saccharomyces cerevisiae Proteins
Ribosomal Proteins
RNA, Small Interfering
Gene Silencing
Protein Folding
Benzodioxoles
Protein Stability
Gene Knockdown Techniques
High-Throughput Screening Assays
Rights: Copyright: © 2016 Veit et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
DOI: 10.1371/journal.pbio.1002462
Appears in Collections:Aurora harvest 3
Molecular and Biomedical Science publications

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