Yiping Yang, Daniel C. Devor, John F. Engelhardt, Stephen A. Ernst, Theresa V. Strong, Francis S. Collins, Jonathan A. Cohn, Raymond A. Frizzell, James M. Wilson

Cystic fibrosis (CF) is caused by mutations in the gene encoding a chloride channel called the CF transmembrane conductance regulator (CFTR). A single mutation in this gene, deletion of three nucleotides that leads to the absence of phenylalanine 508 (i.e., delta F508), is found on 70% of all CF chromosomes. To explore the molecular mechanism(s) responsible for defective chloride transport in patients with CF, we have studied the processing, localization, and function of wild type (W.T.), delta F508 and G551D CFTR (a G-->D missense mutation at position 551) in retrovirus transduced L cells. Cell transduced with W.T. CFTR expressed a 170 kd CFTR protein that was endoglycosidase H (Endo H) resistant, localized to the plasma membrane, and generated a cAMP-mediated anion conductance (GCl) when stimulated with standard concentrations of forskolin (5 microM), cpt cAMP (400 microM) and IBMX (100 microM). The G551D CFTR was indistinguishable from W.T. CFTR with respect to post-translational processing and localization, but it did not produce a cAMP-activated GCl in response to the standard stimulation cocktail. However, raising the IBMX concentration to 4 mM produced GCl in G551D expressing cells. Cells transduced with delta F508 CFTR expressed an Endo H sensitive CFTR protein (approximately 140 kd) that was found in a cytosolic, perinuclear location. These cells did not respond to the standard cocktail, but approximately 20% of cells increased GCl when the cocktail contained 4 mM IBMX.(ABSTRACT TRUNCATED AT 250 WORDS)