Present address: BioPlanta GmbH, Deutscher Platz 5, D-04103 Leipzig, Germany.
Members of the Arabidopsis AtTPK/KCO family form homomeric vacuolar channels in planta
Article first published online: 19 SEP 2006
The Plant Journal
Volume 48, Issue 2, pages 296–306, October 2006
How to Cite
Voelker, C., Schmidt, D., Mueller-Roeber, B. and Czempinski, K. (2006), Members of the Arabidopsis AtTPK/KCO family form homomeric vacuolar channels in planta. The Plant Journal, 48: 296–306. doi: 10.1111/j.1365-313X.2006.02868.x
- Issue published online: 19 SEP 2006
- Article first published online: 19 SEP 2006
- Received 30 March 2006; revised 16 June 2006; accepted 4 July 2006.
- AtTPK/KCO family;
- expression pattern;
The Arabidopsis thaliana K+ channel family of AtTPK/KCO proteins consists of six members including a ‘single-pore’ (Kir-type) and five ‘tandem-pore’ channels. AtTPK4 is currently the only ion channel of this family for which a function has been demonstrated in planta. The protein is located at the plasma membrane forming a voltage-independent K+ channel that is blocked by extracellular calcium ions. In contrast, AtTPK1 is a tonoplast-localized protein, that establishes a K+-selective, voltage-independent ion channel activated by cytosolic calcium when expressed in a heterologous system, i.e. yeast. Here, we provide evidence that other AtTPK/KCO channel subunits, i.e. AtTPK2, AtTPK3, AtTPK5 and AtKCO3, are also targeted to the vacuolar membrane, opening the possibility that they interact at the target membrane to form heteromeric ion channels. However, when testing the cellular expression patterns of AtTPK/KCO genes we observed distinct expression domains that overlap in only a few tissues of the Arabidopsis plant, making it unlikely that different channel subunits interact to form heteromeric channels. This conclusion was substantiated by in planta expression of combinations of selected tonoplast AtTPK/KCO proteins. Fluorescence resonance energy transfer assays indicate that protein interaction occurs between identical channel subunits (most efficiently between AtTPK1 or AtKCO3) but not between different channel subunits. The finding could be confirmed by bimolecular fluorescence complementation assays. We conclude that tonoplast-located AtTPK/KCO subunits form homomeric ion channels in vivo.