Interaction of the K(+)-channel KAT1 with the coat protein complex II coat component Sec24 depends on a di-acidic endoplasmic reticulum export motif.
The correct functioning of ion channels depends not only on the control of their activity but also on the regulation of the number of channels in the membrane. For example, it has been proposed that the density of the plant K(+)-channel KAT1 may be adjusted by controlling its export from its site of synthesis, the endoplasmic reticulum (ER). Efficient transport of the channel to the plasma membrane was found to depend on a di-acidic ER export signal in the C-terminus of the protein. Studies in yeast and mammals indicate that di-acidic ER export motifs are essential for enrichment of proteins into ER-derived coat protein complex II (COPII) vesicles and are recognized by Sec24 a component of the COPII coat. To investigate whether similar mechanisms also exist in plants we have analysed the interaction of KAT1 with Sec24 in vivo using fluorescence resonance energy transfer (FRET) measurements in Vicia faba guard cells. These measurements revealed a FRET signal between KAT1 and Sec24 fused to the cyan fluorescent protein and the yellow fluorescent protein, respectively, indicating an interaction between KAT1 and Sec24. The FRET signal only occurred in the perinuclear region of the ER and was dependent on the di-acidic ER export motif of KAT1. Together, the results point to a highly conserved mechanism for ER export of KAT1 whereby the channel is recruited into COPII vesicles via binding of the di-acidic motif to Sec24.
Sieben
Christian
C
Institute of Botany, Technical University of Darmstadt, 64287 Darmstadt, Germany.
Mikosch
Melanie
M
Brandizzi
Federica
F
Homann
Ulrike
U
D001693:Biological Transport, Active; D022181:COP-Coated Vesicles / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D010940:Plant Proteins / Q000378:metabolism*; D054046:Plant Stomata / Q000378:metabolism; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D054730:Protein Interaction Domains and Motifs; D021382:Protein Sorting Signals; D033921:Vesicular Transport Proteins / Q000378:metabolism; D031307:Vicia faba / Q000378:metabolism*
D010940:Plant Proteins; D024661:Potassium Channels, Inwardly Rectifying; D021382:Protein Sorting Signals; D033921:Vesicular Transport Proteins