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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Diacidic Motif Is Required for Efficient Transport of the K⁺ Channel KAT1 to the Plasma Membrane¹

Institute of Botany, Darmstadt University of Technology, 64287 Darmstadt, Germany

For a number of mammalian ion channels, trafficking to the plasma membrane was found to be controlled by intrinsic sequence motifs. Among these sequences are diacidic motifs that function as endoplasmic reticulum (ER) export signals. So far it is unclear if similar motifs also exist in plant ion channels. In this study we analyzed the function of four diacidic DXE/DXD motifs of the plant K⁺ channel KAT1. Mutation of the first diacidic DXE motif resulted in a strong reduction of the KAT1 conductance in both guard cell protoplasts and HEK293 cells (human embryonic kidney cells). Confocal fluorescence microscopy of guard cells expressing the mutated KAT1 fused to green fluorescent protein revealed localization of the mutated channel only in intracellular structures around the nucleus. These structures could be identified as part of the ER via coexpression of KAT1 fused to yellow fluorescent protein with an ER-retained protein (HDEL) fused to cyan fluorescent protein. Block of vesicle formation from the ER by overexpression of the small GTP-binding protein Sar1 fixed in its GDP-bound form led to retention of wild-type KAT1 in similar parts of the ER. Mutation of the three other diacidic motifs had no effect. Together, the results demonstrate that one diacidic motif of KAT1 is essential for ER export of the functional channel in both guard cell protoplasts and HEK293 cells. This suggests that trafficking of plant plasma membrane ion channels is controlled via a conserved mechanism.

² These authors contributed equally to the paper.

³ Present address: School of Biomedical Sciences, University of Queensland, St. Lucia, QLD 4072, Australia.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Ulrike Homann (homann-u{at}bio.tu-darmstadt.de).

www.plantphysiol.org/cgi/doi/10.1104/pp.106.087064

^* Corresponding author; e-mail homann-u{at}bio.tu-darmstadt.de; fax 49–6151–164630.

Received July 24, 2006; accepted August 29, 2006; published September 1, 2006.

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