An aspartyl protease defines a novel pathway for export of Toxoplasma proteins into the host cell

Infection by Toxoplasma gondii leads to massive changes to the host cell. Here, we identify a novel host cell effector export pathway that requires the Golgi-resident aspartyl protease 5 (ASP5). We demonstrate that ASP5 cleaves a highly constrained amino acid motif that has similarity to the PEXEL-motif of Plasmodium parasites. We show that ASP5 matures substrates at both the N- and C-terminal ends of proteins and also controls trafficking of effectors without this motif. Furthermore, ASP5 controls establishment of the nanotubular network and is required for the efficient recruitment of host mitochondria to the vacuole. Assessment of host gene expression reveals that the ASP5-dependent pathway influences thousands of the transcriptional changes that Toxoplasma imparts on its host cell. All these changes result in attenuation of virulence of Δasp5 tachyzoites in vivo. This work characterizes the first identified machinery required for export of Toxoplasma effectors into the infected host cell. DOI: http://dx.doi.org/10.7554/eLife.10809.001
eLife digest Toxoplasma gondii is a parasite that is thought to infect over two billion people worldwide. Often these infections cause no noticeable symptoms, but can cause serious illness in people with weakened immune systems. Toxoplasma parasites must enter human cells in order to survive. To dramatically increase their chances of survival, the parasites then deliver specialized proteins into the host cell that disarm the host’s immune defenses. Understanding how these specialized proteins are transported from inside the parasite into the host cell, and how this process can be blocked, may lead to new treatments for these and related parasitic infections. By genetically modifying Toxoplasma parasites to lack a parasite enzyme, Coffey et al. have now discovered that this molecule is required for correctly transporting parasite proteins. This enzyme is called aspartyl protease 5 (ASP5) and is found in the parasite in a structure called the Golgi apparatus, which acts as a main hub for protein transport. ASP5 cuts proteins at a ‘barcode’ that is found in many different types of proteins, priming them for transport out of the parasite and for export into the host cell in some cases. Coffey et al. show that in parasites that lack ASP5, these proteins are no longer cleaved and are not transported correctly, blocking the activities that parasites normally perform to ensure their survival. Therefore, ASP5 plays an important role in transporting a wide range of proteins associated with disease, including transporting certain proteins directly into the host cell. Future studies that compare parasites that lack ASP5 to normal parasites will aim to identify new proteins used by the parasites to defeat the host’s immune defenses. DOI: http://dx.doi.org/10.7554/eLife.10809.002