A minimal cysteine motif required to activate the SKOR K+ channel of Arabidopsis by the reactive oxygen species H2O2.
Reactive oxygen species (ROS) are essential for development and stress signaling in plants. They contribute to plant defense against pathogens, regulate stomatal transpiration, and influence nutrient uptake and partitioning. Although both Ca(2+) and K(+) channels of plants are known to be affected, virtually nothing is known of the targets for ROS at a molecular level. Here we report that a single cysteine (Cys) residue within the Kv-like SKOR K(+) channel of Arabidopsis thaliana is essential for channel sensitivity to the ROS H(2)O(2). We show that H(2)O(2) rapidly enhanced current amplitude and activation kinetics of heterologously expressed SKOR, and the effects were reversed by the reducing agent dithiothreitol (DTT). Both H(2)O(2) and DTT were active at the outer face of the membrane and current enhancement was strongly dependent on membrane depolarization, consistent with a H(2)O(2)-sensitive site on the SKOR protein that is exposed to the outside when the channel is in the open conformation. Cys substitutions identified a single residue, Cys(168) located within the S3 α-helix of the voltage sensor complex, to be essential for sensitivity to H(2)O(2). The same Cys residue was a primary determinant for current block by covalent Cys S-methioylation with aqueous methanethiosulfonates. These, and additional data identify Cys(168) as a critical target for H(2)O(2), and implicate ROS-mediated control of the K(+) channel in regulating mineral nutrient partitioning within the plant.
Garcia-Mata
Carlos
C
Laboratory of Plant Physiology and Biophysics, Faculty of Biomedical and Life Sciences, Plant Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
Wang
Jianwen
J
Gajdanowicz
Pawel
P
Gonzalez
Wendy
W
Hills
Adrian
A
Donald
Naomi
N
Riedelsberger
Janin
J
Amtmann
Anna
A
Dreyer
Ingo
I
Blatt
Michael R
MR
D020816:Amino Acid Motifs / Q000235:genetics / Q000502:physiology; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004594:Electrophysiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D056004:Molecular Dynamics Simulation; D018520:Plant Shoots / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D051637:Shaker Superfamily of Potassium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D029681:Arabidopsis Proteins; C114544:SKOR protein, Arabidopsis; D051637:Shaker Superfamily of Potassium Channels; D006861:Hydrogen Peroxide; D011188:Potassium