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The phylum Apicomplexa comprises a group of obligate intracellular parasites of broad medical and agricultural significance, including  Toxoplasma gondii  and the malaria-causing  Plasmodium  spp. Key to their parasitic lifestyle is the need to egress from an infected cell, actively move through tissue, and reinvade another cell, thus perpetuating infection. Ca 2+ -mediated signaling events modulate key steps required for host cell egress, invasion and motility, including secretion of microneme organelles and activation of the force-generating actomyosin-based motor. Here we show that a plant-like Calcium-Dependent Protein Kinase (CDPK) in  T. gondii ,  Tg CDPK3, which localizes to the inner side of the plasma membrane, is not essential to the parasite but is required for optimal  in vitro  growth. We demonstrate that  Tg CDPK3, the orthologue of  Plasmodium Pf CDPK1, regulates Ca 2+  ionophore- and DTT-induced host cell egress, but not motility or invasion. Furthermore, we show that targeting to the inner side of the plasma membrane by dual acylation is required for its activity. Interestingly,  Tg CDPK3 regulates microneme secretion when parasites are intracellular but not extracellular. Indeed, the requirement for  Tg CDPK3 is most likely determined by the high K +  concentration of the host cell. Our results therefore suggest that  Tg CDPK3's role differs from that previously hypothesized, and rather support a model where this kinase plays a role in rapidly responding to Ca 2+  signaling in specific ionic environments to upregulate multiple processes required for gliding motility.

TgCDPK3 Regulates Calcium-Dependent Egress of Toxoplasma gondii from Host Cells