KCNC3R420H, a K+ channel mutation causative in spinocerebellar ataxia 13 displays aberrant intracellular trafficking

•KCNC3R420H displays aberrant glycosylation.•KCNC3R420H is retained in the Golgi by immunohistochemistry and FRET.•Coexpression of KCNC3R420H and KCNC3WT causes retention of both proteins in the Golgi.•KCNC3R420H expression results in Golgi and cell dysmorphology.

Spinocerebellar ataxia 13 (SCA13) is an autosomal dominant disease resulting from mutations in KCNC3 (Kv3.3), a voltage-gated potassium channel. The KCNC3R420H mutation was first identified as causative for SCA13 in a four-generation Filipino kindred with over 20 affected individuals. Electrophysiological analyses in oocytes previously showed that this mutation did not lead to a functional channel and displayed a dominant negative phenotype. In an effort to identify the molecular basis of this allelic form of SCA13, we first determined that human KCNC3WT and KCNC3R420H display disparate post-translational modifications, and the mutant protein has reduced complex glycan adducts. Immunohistochemical analyses demonstrated that KCNC3R420H was not properly trafficking to the plasma membrane and surface biotinylation demonstrated that KCNC3R420H exhibited only 24% as much surface expression as KCNC3WT. KCNC3R420H trafficked through the ER but was retained in the Golgi. KCNC3R420H expression results in altered Golgi and cellular morphology. Electron microscopy of KCNC3R420H localization further supports retention in the Golgi. These results are specific to the KCNC3R420H allele and provide new insight into the molecular basis of disease manifestation in SCA13.