Correcting the F508del-CFTR variant by modulating eukaryotic translation initiation factor 3-mediated translation initiation

J Biol Chem. 2018 Aug 31;293(35):13477-13495. doi: 10.1074/jbc.RA118.003192. Epub 2018 Jul 13.


Inherited and somatic rare diseases result from >200,000 genetic variants leading to loss- or gain-of-toxic function, often caused by protein misfolding. Many of these misfolded variants fail to properly interact with other proteins. Understanding the link between factors mediating the transcription, translation, and protein folding of these disease-associated variants remains a major challenge in cell biology. Herein, we utilized the cystic fibrosis transmembrane conductance regulator (CFTR) protein as a model and performed a proteomics-based high-throughput screen (HTS) to identify pathways and components affecting the folding and function of the most common cystic fibrosis-associated mutation, the F508del variant of CFTR. Using a shortest-path algorithm we developed, we mapped HTS hits to the CFTR interactome to provide functional context to the targets and identified the eukaryotic translation initiation factor 3a (eIF3a) as a central hub for the biogenesis of CFTR. Of note, siRNA-mediated silencing of eIF3a reduced the polysome-to-monosome ratio in F508del-expressing cells, which, in turn, decreased the translation of CFTR variants, leading to increased CFTR stability, trafficking, and function at the cell surface. This finding suggested that eIF3a is involved in mediating the impact of genetic variations in CFTR on the folding of this protein. We posit that the number of ribosomes on a CFTR mRNA transcript is inversely correlated with the stability of the translated polypeptide. Polysome-based translation challenges the capacity of the proteostasis environment to balance message fidelity with protein folding, leading to disease. We suggest that this deficit can be corrected through control of translation initiation.

Keywords: F508del-CFTR; cystic fibrosis; cystic fibrosis transmembrane conductance regulator (CFTR); eIF3a; eukaryotic translation initiation; eukaryotic translation initiation factor 3; network hub; protein misfolding; protein translation; proteostasis; translation initiation factor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cystic Fibrosis Transmembrane Conductance Regulator / chemistry
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism*
  • Eukaryotic Initiation Factor-3 / genetics
  • Eukaryotic Initiation Factor-3 / metabolism*
  • Humans
  • Mutation
  • Peptide Chain Initiation, Translational*
  • Phenylalanine / chemistry
  • Phenylalanine / genetics
  • Phenylalanine / metabolism
  • Protein Folding
  • Protein Interaction Maps
  • Protein Transport
  • RNA Interference
  • RNA, Small Interfering / genetics


  • CFTR protein, human
  • EIF3A protein, human
  • Eukaryotic Initiation Factor-3
  • RNA, Small Interfering
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Phenylalanine