[Mitogen activated protein kinases (MAPK) and Toxoplasma gondii host cell invasion]

Toxoplasmosis is still a big concern in Public Health in France, regarding two particular aspects: congenital toxoplasmosis and reactivated toxoplasmosis in immunodeficient patients. Toxoplasma gondii is an obligate intracellular parasite, that can invade almost all nucleated cells. The invasion step has been widely studied, but its accurate mechanism and the cell receptor remain largely unknown. In this work, we have attempted to investigate the role of kinases and signal transduction in the host cell penetration. We characterized mitogen-activated protein kinases (MAPK). in the parasite itself by immunoblotting, immunofluorescence, and determination of enzymatic activity. In particular, we identified two proteins of 43 and 47 kDa, that could be homologues of extracellular signal-regulated kinases (ERK), i.e. ERK2 and ERK1, respectively. These parasite MAPK are activated by calcium and phorbolmyristyl acetate and inhibited by RO 31-8220, suggesting an activation through protein kinases C (PKC). Indeed the MAPK kinase (= MEK) inhibitor PD98059 inhibits the activation of parasite MAPK, suggesting that a MEK homologue could be responsible for the dual phosphorylation of MAPK on tyrosine and threonine residues, necessary for their activation. Finally, we demonstrated that activation or inhibition of parasite MAPK by preincubation of parasites with various activators or inhibitors led to an increased or reduced host cell invasion in vitro by parasites, respectively. All these results are in favour of a role of ERK-type parasite kinases in T. gondii infectivity in vitro.