Ca2+ entry at the plasma membrane and uptake by acidic stores is regulated by the activity of the V‐H+‐ATPase in Toxoplasma gondii

Ca²⁺ is a universal intracellular signal that regulates many cellular functions. In Toxoplasma gondii, the controlled influx of extracellular and intracellular Ca²⁺ into the cytosol initiates a signaling cascade that promotes pathogenic processes like tissue destruction and dissemination. In this work we studied the role of proton transport in cytosolic Ca²⁺ homeostasis and the initiation of Ca²⁺ signaling. We used a T. gondii mutant of the V-ATPase, a pump previously shown to transport protons to the extracellular medium, control intracellular pH and membrane potential and we show that proton gradients are important for maintaining resting cytosolic Ca²⁺ at physiological levels and for Ca²⁺ influx. Proton transport was also important for Ca²⁺ storage by acidic stores and, unexpectedly, the endoplasmic reticulum. Proton transport impacted the amount of polyphosphate (polyP), a phosphate polymer that binds Ca²⁺ and concentrate in acidocalcisomes. This was supported by the co-localization of the vacuolar transporter chaperone 4 (VTC4), the catalytic subunit of the VTC complex that synthesizes polyP, with the V-ATPase in acidocalcisomes. Our work show that proton transport regulate plasma membrane Ca²⁺ transport and control acidocalcisome polyP and Ca²⁺ content impacting Ca²⁺ signaling and downstream stimulation of motility and egress in T. gondii.

Andrew J Stasic, Eric J Dykes, Ciro D Cordeiro, Stephen A Vella, Mojtaba S Fazli, Shannon Quinn, Roberto Docampo, Silvia N J Moreno. Mol Microbiol. 2021 Apr 1. doi: 10.1111/mmi.14722