title: Functional activation of the cystic fibrosis trafficking mutant delta F508-CFTR by overexpression.
abstract: The most common mutation in the gene associated with cystic fibrosis (CF) causes deletion of phenylalanine at residue 508 (delta F508) of the gene product called CFTR. This mutation results in the synthesis of a variant CFTR protein that is defective in its ability to traffic to the plasma membrane. Because earlier studies showed delta F508-CFTR retains significant phosphorylation-regulated chloride (Cl-) channel activity, processes capable of restoring the mislocalized delta F508-CFTR to the correct cellular destination may have therapeutic benefit. Here we report one such process that involves overexpression of the mutant protein and appears to result in the escape of a small amount of delta F508-CFTR to the plasma membrane. In recombinant cells where expression of delta F508-CFTR is controlled by the metallothionein promoter, this effect can be brought about by treatment with sodium butyrate. Although cAMP-activated Cl- channel activity could also be detected in immortalized human airway epithelial cells homozygous for the delta F508 mutation at the single cell level, treatment with butyrate did not generate a measurable cAMP-stimulated Cl- current in polarized monolayers of primary CF airway epithelia. However, the observation that overexpression can effect the presence of recombinant delta F508-CFTR at the plasma membrane suggests that perhaps other butyrate-like compounds that are more potent and more specific for the promoter of the CF gene may be efficacious in alleviating the Cl- channel defect associated with CF.
authors.affiliation: Genzyme Corporation, Framingham, Massachusetts 01701, USA.
authors.forename: S H
authors.initials: SH
authors.lastname: Cheng
authors.forename: S L
authors.initials: SL
authors.lastname: Fang
authors.forename: J
authors.initials: J
authors.lastname: Zabner
authors.forename: J
authors.initials: J
authors.lastname: Marshall
authors.forename: S
authors.initials: S
authors.lastname: Piraino
authors.forename: S C
authors.initials: SC
authors.lastname: Schiavi
authors.forename: D M
authors.initials: DM
authors.lastname: Jefferson
authors.forename: M J
authors.initials: MJ
authors.lastname: Welsh
authors.forename: A E
authors.initials: AE
authors.lastname: Smith
mesh_terms: D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D018118:Chloride Channels / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D000242:Cyclic AMP / Q000502:physiology; D003550:Cystic Fibrosis / Q000378:metabolism / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005456:Fluorescent Dyes; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D010179:Pancreas / Q000378:metabolism / Q000473:pathology; D011401:Promoter Regions, Genetic; D011806:Quinolinium Compounds; D011994:Recombinant Proteins; D012137:Respiratory System / Q000166:cytology / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D002087:Butyrates; C505032:CFTR protein, human; D018118:Chloride Channels; D002712:Chlorides; D005456:Fluorescent Dyes; D011806:Quinolinium Compounds; D011994:Recombinant Proteins; C093349:cystic fibrosis transmembrane conductance regulator delta F508; D020148:Butyric Acid; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C052335:6-methoxy-N-(3-sulfopropyl)quinolinium; D008668:Metallothionein; D000242:Cyclic AMP