Processing and secretion of ROP13: A unique Toxoplasma effector protein.
Like most intracellular pathogens, Toxoplasma synthesizes and secretes an arsenal of proteins to successfully invade its host cell and hijack host functions for intracellular survival. The rhoptries are key secretory organelles that inject proteins into the host cell where they are positioned to co-opt host processes, although little is known regarding how these proteins exert their functions. We show here that the rhoptry protein ROP13 is synthesized as a pre-pro-protein that is processed in the parasite. Processing occurs at a conserved SphiXE cleavage site as mutagenesis of glutamic acid to alanine at the P1 position disrupts ROP13 maturation. We also demonstrate that processing of the prodomain is not necessary for rhoptry targeting and secretion. While gene disruption reveals that ROP13 is not essential for growth in fibroblasts in vitro or for virulence in vivo, we find that ROP13 is a soluble effector protein that can access the cytoplasm of host cells. Exogenously expressed ROP13 in human cells remains cytosolic but also appears toxic, suggesting that over-expression of this effector protein is disrupting some function within the host cell.
Turetzky
Jay M
JM
Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095-1489, USA.
Chu
David K
DK
Hajagos
Bettina E
BE
Bradley
Peter J
PJ
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000469:parasitology; D055786:Gene Knockout Techniques; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D011499:Protein Processing, Post-Translational; D015800:Protozoan Proteins / Q000378:metabolism*; D014122:Toxoplasma / Q000378:metabolism* / Q000472:pathogenicity; D014123:Toxoplasmosis / Q000469:parasitology / Q000473:pathology; D014774:Virulence; D037521:Virulence Factors / Q000378:metabolism*
D015800:Protozoan Proteins; D037521:Virulence Factors