The arginine-rich N-terminal domain of ROP18 is necessary for vacuole targeting and virulence of Toxoplasma gondii.
Toxoplasma gondii uses specialized secretory organelles called rhoptries to deliver virulence determinants into the host cell during parasite invasion. One such determinant called rhoptry protein 18 (ROP18) is a polymorphic serine/threonine kinase that phosphorylates host targets to modulate acute virulence. Following secretion into the host cell, ROP18 traffics to the parasitophorous vacuole membrane (PVM) where it is tethered to the cytosolic face of this host-pathogen interface. However, the functional consequences of PVM association are not known. In this report, we show that ROP18 mutants altered in an arginine-rich domain upstream of the kinase domain fail to associate to the PVM following secretion from rhoptries. During infection, host cells upregulate immunity-related GTPases that localize to and destroy the PVM surrounding the parasites. ROP18 disarms this host innate immune pathway by phosphorylating IRGs in a critical GTPase domain and preventing loading on the PVM. Vacuole-targeting mutants of ROP18 failed to phosphorylate Irga6 and were unable to divert IRGs from the PVM, despite retaining intrinsic kinase activity. As a consequence, these mutants were avirulent in a mouse model of acute toxoplasmosis. Thus, the association of ROP18 with the PVM, mediated by its N-terminal arginine-rich domain, is critical to its function as a virulence determinant.
Department of Molecular Microbiology, Washington University School of Medicine, St Louis, MO, USA.
Sarah J
SJ
Fentress
Tobias
T
Steinfeldt
Jonathan C
JC
Howard
L David
LD
Sibley
D000818:Animals; D004195:Disease Models, Animal; D051379:Mice; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D015800:Protozoan Proteins; D017384:Sequence Deletion; D014122:Toxoplasma / Q000472:pathogenicity*; D014124:Toxoplasmosis, Animal / Q000469:parasitology / Q000473:pathology; D014617:Vacuoles / Q000378:metabolism* / Q000469:parasitology*; D014774:Virulence; D037521:Virulence Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
D015800:Protozoan Proteins; D037521:Virulence Factors; D017346:Protein Serine-Threonine Kinases; C556793:ROP18 protein, Toxoplasma gondii