title: Lotus japonicus CASTOR and POLLUX are ion channels essential for perinuclear calcium spiking in legume root endosymbiosis.
abstract: The mechanism underlying perinuclear calcium spiking induced during legume root endosymbioses is largely unknown. Lotus japonicus symbiosis-defective castor and pollux mutants are impaired in perinuclear calcium spiking. Homology modeling suggested that the related proteins CASTOR and POLLUX might be ion channels. Here, we show that CASTOR and POLLUX form two independent homocomplexes in planta. CASTOR reconstituted in planar lipid bilayers exhibited ion channel activity, and the channel characteristics were altered in a symbiosis-defective mutant carrying an amino acid replacement close to the selectivity filter. Permeability ratio determination and competition experiments reveled a weak preference of CASTOR for cations such as potassium over anions. POLLUX has an identical selectivity filter region and complemented a potassium transport-deficient yeast mutant, suggesting that POLLUX is also a potassium-permeable channel. Immunogold labeling localized the endogenous CASTOR protein to the nuclear envelope of Lotus root cells. Our data are consistent with a role of CASTOR and POLLUX in modulating the nuclear envelope membrane potential. They could either trigger the opening of calcium release channels or compensate the charge release during the calcium efflux as counter ion channels.
authors.affiliation: Ludwig-Maximilians-Universität München, Faculty of Biology, Genetics, 80638 München, Germany.
authors.forename: Myriam
authors.initials: M
authors.lastname: Charpentier
authors.forename: Rolf
authors.initials: R
authors.lastname: Bredemeier
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wanner
authors.forename: Naoya
authors.initials: N
authors.lastname: Takeda
authors.forename: Enrico
authors.initials: E
authors.lastname: Schleiff
authors.forename: Martin
authors.initials: M
authors.lastname: Parniske
mesh_terms: D020013:Calcium Signaling / Q000235:genetics / Q000502:physiology*; D005816:Genetic Complementation Test; D015151:Immunoblotting; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D000070116:Lotus / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D013559:Symbiosis / Q000235:genetics / Q000502:physiology*; D020798:Two-Hybrid System Techniques
chemical_list: D007473:Ion Channels; D010940:Plant Proteins