Functional Analysis of Missense Mutations in Kv8.2 Causing Cone Dystrophy with Supernormal Rod Electroretinogram

Bradley J. Jarvis,Susan E. Wilkie,David M. Hunt,Martin Stocker,Katie E. Smith,Joseph T. Tebbs-Warner,Linn Gallasch

doi:10.1074/jbc.m112.388033

Bradley J. Jarvis, Susan E. Wilkie + Show 5 more

Open Access

https://doi.org/10.1074/jbc.m112.388033

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Abstract

Mutations in KCNV2 have been proposed as the molecular basis for cone dystrophy with supernormal rod electroretinogram. KCNV2 codes for the modulatory voltage-gated potassium channel α-subunit, Kv8.2, which is incapable of forming functional channels on its own. Functional heteromeric channels are however formed with Kv2.1 in heterologous expression systems, with both α-subunit genes expressed in rod and cone photoreceptors. Of the 30 mutations identified in the KCNV2 gene, we have selected three missense mutations localized in the potassium channel pore and two missense mutations localized in the tetramerization domain for analysis. We characterized the differences between homomeric Kv2.1 and heteromeric Kv2.1/Kv8.2 channels and investigated the influence of the selected mutations on the function of heteromeric channels. We found that two pore mutations (W467G and G478R) led to the formation of nonconducting heteromeric Kv2.1/Kv8.2 channels, whereas the mutations localized in the tetramerization domain prevented heteromer generation and resulted in the formation of homomeric Kv2.1 channels only. Consequently, our study suggests the existence of two distinct molecular mechanisms involved in the disease pathology.

Highlights

Homozygosity mapping linked mutations in KCNV2, encoding Kv8.2, to an inherited retinal disorder
Of the 30 mutations identified in the KCNV2 gene, we have selected three missense mutations localized in the potassium channel pore and two missense mutations localized in the tetramerization domain for analysis
We found that two pore mutations (W467G and G478R) led to the formation of nonconducting heteromeric Kv2.1/Kv8.2 channels, whereas the mutations localized in the tetramerization domain prevented heteromer generation and resulted in the formation of homomeric Kv2.1 channels only

Summary

Introduction

Homozygosity mapping linked mutations in KCNV2, encoding Kv8.2, to an inherited retinal disorder. Results: Mutant Kv8.2 subunits render heteromeric Kv2.1/Kv8.2 channels nonfunctional or fail to form heteromers, resulting in homomeric Kv2.1 channels. Conclusion: clinically indistinguishable, different missense mutations impair channel function via two distinct molecular mechanisms. Functional heteromeric channels are formed with Kv2.1 in heterologous expression systems, with both ␣-subunit genes expressed in rod and cone photoreceptors. Of the 30 mutations identified in the KCNV2 gene, we have selected three missense mutations localized in the potassium channel pore and two missense mutations localized in the tetramerization domain for analysis. We found that two pore mutations (W467G and G478R) led to the formation of nonconducting heteromeric Kv2.1/Kv8.2 channels, whereas the mutations localized in the tetramerization domain prevented heteromer generation and resulted in the formation of homomeric Kv2.1 channels only. Our study suggests the existence of two distinct molecular mechanisms involved in the disease pathology

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Conclusion