A novel KCNJ2 nonsense mutation, S369X, impedes trafficking and causes a limited form of Andersen-Tawil syndrome

Circ Cardiovasc Genet. 2011 Jun;4(3):253-60. doi: 10.1161/CIRCGENETICS.110.958157. Epub 2011 Apr 14.

Abstract

Background: Mutations in KCNJ2, a gene encoding the inward rectifier K(+) channel Kir2.1, are associated with Andersen-Tawil syndrome (ATS), which is characterized by (1) ventricular tachyarrhythmias associated with QT (QU)-interval prolongation, (2) periodic paralysis, and (3) dysmorphic features.

Methods and results: We identified a novel KCNJ2 mutation, S369X, in a 13-year-old boy with prominent QU-interval prolongation and mild periodic paralysis. The mutation results in the truncation at the middle of the cytoplasmic C-terminal domain that eliminates the endoplasmic reticulum (ER)-to-Golgi export signal. Current recordings from Chinese hamster ovary cells transfected with KCNJ2-S369X exhibited significantly smaller K(+) currents compared with KCNJ2 wild type (WT) (1 μg each) (-84 ± 14 versus -542 ± 46 picoamperes per picofarad [pA/pF]; -140 mV; P<0.0001). Coexpression of the WT and S369X subunits did not show a dominant-negative suppression effect but yielded larger currents than those of WT+S369X (-724 ± 98 pA/pF>-[84+542] pA/pF; 1 μg each; -140 mV). Confocal microscopy analysis showed that the fluorescent protein-tagged S369X subunits were predominantly retained in the ER when expressed alone; however, the expression of S369X subunits to the plasma membrane was partially restored when coexpressed with WT. Fluorescence resonance energy transfer analysis demonstrated direct protein-protein interactions between WT and S369X subunits in the intracellular compartment.

Conclusions: The S369X mutation causes a loss of the ER export motif. However, the trafficking deficiency can be partially rescued by directly assembling with the WT protein, resulting in a limited restoration of plasma membrane localization and channel function. This alleviation may explain why our patient presented with a relatively mild ATS phenotype.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Amino Acid Sequence
  • Andersen Syndrome / genetics*
  • Andersen Syndrome / physiopathology
  • Animals
  • CHO Cells
  • COS Cells
  • Chlorocebus aethiops
  • Codon, Nonsense*
  • Cricetinae
  • Cricetulus
  • Electrocardiography
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Male
  • Molecular Sequence Data
  • Patch-Clamp Techniques
  • Pedigree
  • Potassium Channels, Inwardly Rectifying / genetics*
  • Potassium Channels, Inwardly Rectifying / metabolism
  • Protein Transport*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Alignment

Substances

  • Codon, Nonsense
  • KCNJ2 protein, human
  • Potassium Channels, Inwardly Rectifying
  • Recombinant Fusion Proteins