Vicia faba SV channel VfTPC1 is a hyperexcitable variant of plant vacuole Two Pore Channels

Abstract

To fire action-potential-like electrical signals, the vacuole membrane requires the two-pore channel TPC1, formerly called SV channel. The TPC1/SV channel functions as a depolarization-stimulated, non-selective cation channel that is inhibited by luminal Ca²⁺. In our search for species-dependent functional TPC1 channel variants with different luminal Ca²⁺ sensitivity, we found in total three acidic residues present in Ca²⁺ sensor sites 2 and 3 of the Ca²⁺-sensitive AtTPC1 channel from Arabidopsis thaliana that were neutral in its Vicia faba ortholog and also in those of many other Fabaceae. When expressed in the Arabidopsis AtTPC1-loss-of-function background, wild-type VfTPC1 was hypersensitive to vacuole depolarization and only weakly sensitive to blocking luminal Ca²⁺. When AtTPC1 was mutated for these VfTPC1-homologous polymorphic residues, two neutral substitutions in Ca²⁺ sensor site 3 alone were already sufficient for the Arabidopsis At-VfTPC1 channel mutant to gain VfTPC1-like voltage and luminal Ca²⁺ sensitivity that together rendered vacuoles hyperexcitable. Thus, natural TPC1 channel variants exist in plant families which may fine-tune vacuole excitability and adapt it to environmental settings of the particular ecological niche.

Keywords: A. thaliana; Brassicaceae; Fabaceae; molecular biophysics; plant biology; pore; potassium channel; structural biology; vacuolar calcium sensor; voltage gating.