An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca²⁺ signaling and motility in Toxoplasma gondii.

ABSTRACT: Protozoan parasites of the phylum Apicomplexa actively move through tissue to initiate and perpetuate infection. The regulation of parasite motility relies on cyclic nucleotide-dependent kinases, but how these kinases are activated remains unknown. Here, using an array of biochemical and cell biology approaches, we show that the apicomplexan parasite Toxoplasma gondii expresses a large guanylate cyclase (TgGC) protein, which contains several upstream ATPase transporter-like domains. We show that TgGC has a dynamic localization, being concentrated at the apical tip in extracellular parasites, which then relocates to a more cytosolic distribution during intracellular replication. Conditional TgGC knockdown revealed that this protein is essential for acute-stage tachyzoite growth, as TgGC-deficient parasites were defective in motility, host cell attachment, invasion, and subsequent host cell egress. We show that TgGC is critical for a rapid rise in cytosolic [Ca²⁺] and for secretion of microneme organelles upon stimulation with a cGMP agonist, but these deficiencies can be bypassed by direct activation of signaling by a Ca²⁺ ionophore. Furthermore, we found that TgGC is required for transducing changes in extracellular pH and [K⁺] to activate cytosolic [Ca²⁺] flux. Together, the results of our work implicate TgGC as a putative signal transducer that activates Ca²⁺ signaling and motility in Toxoplasma.

SUBMITTER: Yang L 

PROVIDER: S-EPMC6552420 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature