Inhibition of protein kinase CK2 closes the CFTR Cl channel, but has no effect on the cystic fibrosis mutant deltaF508-CFTR

K. J. Treharne, Z. Xu, J. H. Chen, O. G. Best, D. M. Cassidy, D. C. Gruenert, P. Hegyi, M. A. Gray, D. N. Sheppard, K. Kunzelmann, A. Mehta

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

BACKGROUND: Deletion of phenylalanine-508 (DeltaF508) from the first nucleotide-binding domain (NBD1) in the wild-type cystic fibrosis (CF) transmembrane-conductance regulator (wtCFTR) causes CF. However, the mechanistic relationship between DeltaF508-CFTR and the diversity of CF disease is unexplained. The surface location of F508 on NBD1 creates the potential for protein-protein interactions and nearby, lies a consensus sequence (SYDE) reported to control the pleiotropic protein kinase CK2. METHODS: Electrophysiology, immunofluorescence and biochemistry applied to CFTR-expressing cells, Xenopus oocytes, pancreatic ducts and patient biopsies. RESULTS: Irrespective of PKA activation, CK2 inhibition (ducts, oocytes, cells) attenuates CFTR-dependent Cl(-) transport, closing wtCFTR in cell-attached membrane patches. CK2 and wtCFTR co-precipitate and CK2 co-localized with wtCFTR (but not DeltaF508-CFTR) in apical membranes of human airway biopsies. Comparing wild-type and DeltaF508CFTR expressing oocytes, only DeltaF508-CFTR Cl(-) currents were insensitive to two CK2 inhibitors. Furthermore, wtCFTR was inhibited by injecting a peptide mimicking the F508 region, whereas the DeltaF508-equivalent peptide had no effect. CONCLUSIONS: CK2 controls wtCFTR, but not DeltaF508-CFTR. Others find that peptides from the F508 region of NBD1 allosterically control CK2, acting through F508. Hence, disruption of CK2-CFTR interaction by DeltaF508-CFTR might disrupt multiple, membrane-associated, CK2-dependent pathways, creating a new molecular disease paradigm for deleted F508 in CFTR.
Original languageEnglish
Pages (from-to)347-360
Number of pages14
JournalCellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
Volume24
Issue number5-6
DOIs
Publication statusPublished - Nov 2009
Externally publishedYes

Keywords

  • Allosteric Regulation Amino Acid Sequence Animals Biological Transport Casein Kinase II/analysis/antagonists & inhibitors/*metabolism Cell Line Chloride Channels/*metabolism Cricetinae Cyclic AMP/metabolism Cystic Fibrosis/genetics Cystic Fibrosis Transmembrane Conductance Regulator/analysis/genetics/*metabolism Electrophysiological Phenomena Guinea Pigs Humans Immunoprecipitation Molecular Sequence Data Mutation Oocytes/metabolism Protein Interaction Domains and Motifs Xenopus

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