Functional expression of the voltage-gated neuronal mammalian potassium channel rat ether à go-go1 in yeast

It has been shown previously that heterologous expression of inwardly rectifying potassium channels (K⁺-channels) from plants and mammals in K⁺-transport defective yeast mutants can restore the ability of growth in media with low [K⁺]. In this study, the functional expression of an outward rectifyin...

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Bibliographic Details
Published in:FEMS yeast research 2008-05, Vol.8 (3), p.405-413
Main Authors: Schwarzer, Sarah, Kolacna, Lucie, Lichtenberg-Fraté, Hella, Sychrova, Hana, Ludwig, Jost
Format: Article
Language:English
Subjects:
Animals
Barium Compounds - pharmacology
Chlorides - pharmacology
Ether-A-Go-Go Potassium Channels - genetics
Ether-A-Go-Go Potassium Channels - physiology
Hydrogen-Ion Concentration
Mammals
Metal ions
Mutants
N-Terminus
Phenotypes
Potassium
Potassium - metabolism
Potassium Channel Blockers - pharmacology
Potassium channels
Potassium channels (inwardly-rectifying)
Potassium channels (voltage-gated)
Quinidine
Rats
Recombinant Proteins - metabolism
Rubidium - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Yeast
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Summary:It has been shown previously that heterologous expression of inwardly rectifying potassium channels (K⁺-channels) from plants and mammals in K⁺-transport defective yeast mutants can restore the ability of growth in media with low [K⁺]. In this study, the functional expression of an outward rectifying mammalian K⁺-channel in yeast is presented for the first time. The outward-rectifying mammalian neuronal K⁺-channel rat ether à go-go channel 1 (rEAG1, Kv 10.1) was expressed in yeast (Saccharomyces cerevisiae) strains lacking the endogenous K⁺-uptake systems and/or alkali-metal-cation efflux systems. It was found that a truncated channel version, lacking almost the complete intracellular N-terminus (rEAG1Δ190) but not the full-length rEAG1, partially complemented the growth defect of K⁺-uptake mutant cells (trk1,2Δ tok1Δ) in media containing low K⁺ concentrations. The expression of rEAG1Δ190 in a strain lacking the cation efflux systems (nha1Δ ena1-4Δ) increased the sensitivity to high monovalent cation concentrations. Both phenotypes were observed, when rEAG1Δ190 was expressed in a trk1,2Δ and nha1, ena1-4Δ mutant strain. In the presence of K⁺-channel blockers (Cs⁺, Ba²⁺ and quinidine), the growth advantage of rEAG1Δ190 expressing trk1,2 tok1Δ cells disappeared, indicating its dependence on functional rEAG1 channels. The results demonstrate that S. cerevisiae is a suitable expression system even for voltage-gated outward-rectifying mammalian K⁺-channels.
ISSN:1567-1356
1567-1364
DOI:10.1111/j.1567-1364.2007.00351.x