title: Loss of the vacuolar cation channel, AtTPC1, does not impair Ca2+ signals induced by abiotic and biotic stresses.
abstract: The putative two-pore Ca(2+) channel TPC1 has been suggested to be involved in responses to abiotic and biotic stresses. We show that AtTPC1 co-localizes with the K(+)-selective channel AtTPK1 in the vacuolar membrane. Loss of AtTPC1 abolished Ca(2+)-activated slow vacuolar (SV) currents, which were increased in AtTPC1-over-expressing Arabidopsis compared to the wild-type. A Ca(2+)-insensitive vacuolar cation channel, as yet uncharacterized, could be resolved in tpc1-2 knockout plants. The kinetics of ABA- and CO(2)-induced stomatal closure were similar in wild-type and tpc1-2 knockout plants, excluding a role of SV channels in guard-cell signalling in response to these physiological stimuli. ABA-, K(+)-, and Ca(2+)-dependent root growth phenotypes were not changed in tpc1-2 compared to wild-type plants. Given the permeability of SV channels to mono- and divalent cations, the question arises as to whether TPC1 in vivo represents a pathway for Ca(2+) entry into the cytosol. Ca(2+) responses as measured in aequorin-expressing wild-type, tpc1-2 knockout and TPC1-over-expressing plants disprove a contribution of TPC1 to any of the stimulus-induced Ca(2+) signals tested, including abiotic stresses (cold, hyperosmotic, salt and oxidative), elevation in extracellular Ca(2+) concentration and biotic factors (elf18, flg22). In good agreement, stimulus- and Ca(2+)-dependent gene activation was not affected by alterations in TPC1 expression. Together with our finding that the loss of TPC1 did not change the activity of hyperpolarization-activated Ca(2+)-permeable channels in the plasma membrane, we conclude that TPC1, under physiological conditions, functions as a vacuolar cation channel without a major impact on cytosolic Ca(2+) homeostasis.
authors.affiliation: Leibniz Institute of Plant Biochemistry, Stress and Developmental Biology, Weinberg 3, D-06120 Halle, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Ranf
authors.forename: Petra
authors.initials: P
authors.lastname: Wünnenberg
authors.forename: Justin
authors.initials: J
authors.lastname: Lee
authors.forename: Dirk
authors.initials: D
authors.lastname: Becker
authors.forename: Marcel
authors.initials: M
authors.lastname: Dunkel
authors.forename: Rainer
authors.initials: R
authors.lastname: Hedrich
authors.forename: Dierk
authors.initials: D
authors.lastname: Scheel
authors.forename: Petra
authors.initials: P
authors.lastname: Dietrich
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D020013:Calcium Signaling; D002412:Cations / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D009154:Mutation; D018408:Patch-Clamp Techniques; D018515:Plant Leaves / Q000378:metabolism; D018517:Plant Roots / Q000254:growth & development; D015221:Potassium Channels / Q000378:metabolism; D015533:Transcriptional Activation; D014617:Vacuoles / Q000378:metabolism*; D000097903:Two-Pore Channels
chemical_list: D000040:Abscisic Acid; D029681:Arabidopsis Proteins; D002118:Calcium; D015220:Calcium Channels; D002412:Cations; D015221:Potassium Channels; C498240:TPC1 protein, Arabidopsis; D000097903:Two-Pore Channels