A Signaling Factor Linked to Toxoplasma gondii Guanylate Cyclase Complex Controls Invasion and Egress during Acute and Chronic Infection

Toxoplasma gondii is an intracellular apicomplexan parasite that relies on cyclic GMP (cGMP)-dependent signaling to trigger timely egress from host cells in response to extrinsic and intrinsic signals. A guanylate cyclase (GC) complex, conserved across the Apicomplexa, plays a pivotal role in integrating these signals, such as the key lipid mediator phosphatidic acid and changes in pH and ionic composition. This complex is composed of an atypical GC fused to a flippase-like P4-ATPase domain and assembled with the cell division control protein CDC50.1 and a unique GC organizer (UGO). While the dissemination of the fast-replicating tachyzoites responsible for acute infection is well understood, it is less clear if the cyst-forming bradyzoites can disseminate and contribute to cyst burden. Here, we characterized a novel component of the GC complex recently termed signaling linking factor (SLF). Tachyzoites conditionally depleted in SLF are impaired in microneme exocytosis, conoid extrusion, and motility and hence unable to invade and egress. A stage-specific promoter swap strategy allowed the generation of SLF- and GC-deficient bradyzoites that are viable as tachyzoites but show a reduction in cyst burden during the onset of chronic infection. Upon oral infection, SLF-deficient cysts failed to establish infection in mice, suggesting SLF's importance for the natural route of T. gondii infection. IMPORTANCE Toxoplasma gondii is an obligate intracellular parasite of the phylum Apicomplexa. This life-threatening opportunistic pathogen establishes a chronic infection in human and animals that is resistant to immune attacks and chemotherapeutic intervention. The slow-growing parasites persist in tissue cysts that constitute a predominant source of transmission. Host cell invasion and egress are two critical steps of the parasite lytic cycle that are governed by a guanylate cyclase complex conserved across the Apicomplexa. A signaling linked factor is characterized here as an additional component of the complex that not only is essential during acute infection but also plays a pivotal role during natural oral infection with tissue cysts' dissemination and persistence. Toxoplasma gondii is an obligate intracellular parasite of the phylum Apicomplexa. This life-threatening opportunistic pathogen establishes a chronic infection in human and animals that is resistant to immune attacks and chemotherapeutic intervention.