NHE-RF1 protein rescues ΔF508-CFTR function

Florian Bossard, Amal Robay, Gilles Toumaniantz, Shehrazade Dahimene, Frédéric Becq, Jean Merot, Chantal Gauthier

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

In cystic fibrosis (CF), the ΔF508-CFTR anterograde trafficking from the endoplasmic reticulum to the plasma membrane is inefficient. New strategies for increasing the delivery of ΔF508-CFTR to the apical membranes are thus pathophysiologically relevant targets to study for CF treatment. Recent studies have demonstrated that PDZ-containing proteins play an essential role in determining polarized plasma membrane expression of ionic transporters. In the present study we have hypothesized that the PDZ-containing protein NHE-RF1, which binds to the carboxy terminus of CFTR, rescues ΔF508-CFTR expression in the apical membrane of epithelial cells. The plasmids encoding ΔF508-CFTR and NHE-RF1 were intranuclearly injected in A549 or Madin-Darby canine kidney (MDCK) cells, and ΔF508-CFTR channel activity was functionally assayed using SPQ fluorescent probe. Cells injected with ΔF508-CFTR alone presented a low chloride channel activity, whereas its coexpression with NHE-RF1 significantly increased both the basal and forskolin-activated chloride conductances. This last effect was lost with ΔF508-CFTR deleted of its 13 last amino acids or by injection of a specific NHE-RF1 antisense oligonucleotide, but not by NHE-RF1 sense oligonucleotide. Immunocytochemical analysis performed in MDCK cells transiently transfected with ΔF508-CFTR further revealed that NHE-RF1 specifically determined the apical plasma membrane expression of ΔF508-CFTR but not that of a trafficking defective mutant potassium channel (KCNQ1). These data demonstrate that the modulation of the expression level of CFTR protein partners, like NHE-RF1, can rescue ΔF508-CFTR activity.

Original languageEnglish
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume292
Issue number5
DOIs
Publication statusPublished - May 2007
Externally publishedYes

Fingerprint

Madin Darby Canine Kidney Cells
Cell Membrane
Cystic Fibrosis
KCNQ1 Potassium Channel
Cystic Fibrosis Transmembrane Conductance Regulator
Chloride Channels
Proteins
Membranes
Antisense Oligonucleotides
Colforsin
Fluorescent Dyes
Oligonucleotides
Endoplasmic Reticulum
Chlorides
Plasmids
Epithelial Cells
Amino Acids
Injections
Therapeutics

Keywords

  • ΔF508 cystic fibrosis transmembrane conductance regulator
  • Cystic fibrosis
  • Na/H exchanger regulatory factor isoform 1
  • Polarized expression
  • Traffic

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology (medical)

Cite this

NHE-RF1 protein rescues ΔF508-CFTR function. / Bossard, Florian; Robay, Amal; Toumaniantz, Gilles; Dahimene, Shehrazade; Becq, Frédéric; Merot, Jean; Gauthier, Chantal.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 292, No. 5, 05.2007.

Research output: Contribution to journalArticle

Bossard, Florian ; Robay, Amal ; Toumaniantz, Gilles ; Dahimene, Shehrazade ; Becq, Frédéric ; Merot, Jean ; Gauthier, Chantal. / NHE-RF1 protein rescues ΔF508-CFTR function. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2007 ; Vol. 292, No. 5.
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