Toxoplasma gondii PPM3C, a secreted protein phosphatase, affects parasitophorous vacuole effector export

• Published in: PLoS pathogens 2020-12, Vol.16 (12), p.e1008771-e1008771
• Main Authors: Mayoral, Joshua, Tomita, Tadakimi, Tu, Vincent, Aguilan, Jennifer T, Sidoli, Simone, Weiss, Louis M
• Format: Article
• Language: English
• Subjects: Amino acids; Animals; Biology and Life Sciences; Cellular proteins; Chronic infection; Cysts; Genetic aspects; Genetic regulation; HA protein; Health aspects; Host-Pathogen Interactions - physiology; Human populations; Infections; Kinases; Magnesium; Medicine and Health Sciences; Mice; Parasites; Parasitophorous vacuole; Peptides; Phosphatase; Phosphoprotein Phosphatases - metabolism; Phosphorylation; Protein phosphatase; Protein Transport; Proteins; Stop codon; Toxoplasma; Toxoplasma - enzymology; Toxoplasma gondii; Toxoplasmosis - metabolism; Vacuoles - metabolism; Virulence; Virulence Factors - metabolism
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1008771

The intracellular parasite Toxoplasma gondii infects a large proportion of humans worldwide and can cause adverse complications in the settings of immune-compromise and pregnancy. T. gondii thrives within many different cell types due in part to its residence within a specialized and heavily modified compartment in which the parasite divides, termed the parasitophorous vacuole. Within this vacuole, numerous proteins optimize intracellular survival following their secretion by the parasite. We investigated the contribution of one of these proteins, TgPPM3C, predicted to contain a PP2C-class serine/threonine phosphatase domain and previously shown to interact with the protein MYR1, an essential component of a putative vacuolar translocon that mediates effector export into the host cell. Parasites lacking the TgPPM3C gene exhibit a minor growth defect in vitro, are avirulent during acute infection in mice, and form fewer cysts in mouse brain during chronic infection. Phosphoproteomic assessment of TgPPM3C deleted parasite cultures demonstrated alterations in the phosphorylation status of many secreted vacuolar proteins including two exported effector proteins, GRA16 and GRA28, as well as MYR1. Parasites lacking TgPPM3C are defective in GRA16 and GRA28 export, but not in the export of other MYR1-dependant effectors. Phosphomimetic mutation of two GRA16 serine residues results in export defects, suggesting that de-phosphorylation is a critical step in the process of GRA16 export. These findings provide another example of the emerging role of phosphatases in regulating the complex environment of the T. gondii parasitophorous vacuole and influencing the export of specific effector proteins from the vacuolar lumen into the host cell.