Correctors of mutant CFTR enhance subcortical cAMP–PKA signaling through modulating ezrin phosphorylation and cytoskeleton organization

Journal article

The most common mutation of the cystic fibrosis transmembrane regulator (CFTR) gene, F508del, produces a misfolded protein resulting in its defective trafficking to the cell surface and an impaired chloride secretion. Pharmacological treatments partially rescue F508del CFTR activity either directly by interacting with the mutant protein and/or indirectly by altering the cellular protein homeostasis. Here, we show that the phosphorylation of ezrin together with its binding to phosphatidylinositol-4,5-bisphosphate (PIP2) tethers the F508del CFTR to the actin cytoskeleton, stabilizing it on the apical membrane and rescuing the sub-membrane compartmentalization of cAMP and activated PKA. Both the small molecules trimethylangelicin (TMA) and VX-809, which act as ‘correctors’ for F508del CFTR by rescuing F508del-CFTR-dependent chloride secretion, also restore the apical expression of phosphorylated ezrin and actin organization and increase cAMP and activated PKA submembrane compartmentalization in both primary and secondary cystic fibrosis airway cells. Latrunculin B treatment or expression of the inactive ezrin mutant T567A reverse the TMA and VX-809-induced effects highlighting the role of corrector-dependent ezrin activation and actin re-organization in creating the conditions to generate a sub-cortical cAMP pool of adequate amplitude to activate the F508del-CFTR-dependent chloride secretion.

Authors: Abbattiscianni, AC; Favia, M; Mancini, MT; Cardone, RA; Guerra, L

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Company of Biologists
• Journal: Journal of Cell Science
• Volume: 129
• Issue: 6
• Pages: 1128–1140
• Publication date: 2016-03-15
• Acceptance date: 2016-01-21
• DOI: 10.1242/jcs.177907
• EISSN: 1477-9137
• ISSN: 0021-9533
• Language: English
• Keywords: airway cells; cAMP; cystic fibrosis; FRET; PKA; correctors