Figure 8. GRA24 requires ASP5 for export but not processing.

(A) Localization of GRA24-Myc₃ in both WT (Δku80) and Δku80Δasp5 tachyzoites. GRA24 can be observed in the host nucleus and in the PV in WT:GRA24-Myc₃ parasites, whereas this export is lost in the Δasp5:GRA24-Myc₃ parasites. Arrows signify the position of host nuclei (DAPI). (B) The size of GRA24-Myc₃ appears unchanged in the absence of ASP5. αCatalase serves as a loading control. (C) ASP5 cannot cleave peptides containing non-canonical TEXEL-like motifs found within GRA24, compared with cleavage of the GRA16 RRLAE peptide and AAAAE controls. Scale bar is 5 μm. ASP5, Aspartyl Protease 5; DAPI, 4',6-diamidino-2-phenylindole; PV, parasitophorous vacuole; TEXEL, Toxoplasma export element; WT, wild type.

DOI: http://dx.doi.org/10.7554/eLife.10809.015

Figure 8—figure supplement 1. Complementation of Δasp5 parasites restores export of GRA24.

Localisation of transiently transfected GRA24-Myc₃ of in both Δasp5_CRISPR and Δasp5_CRISPR:ASP5_WT-HA₃ (Clone 1) tachyzoites. Filled arrowheads signify the position of host nuclei (DAPI), open arrowheads identify non-transfected parasites and GAP45 marks the periphery of tachyzoites. Scale bar is 5 μm. DAPI, 4',6-diamidino-2-phenylindole; HA₃, triple-hemagglutinin

DOI: http://dx.doi.org/10.7554/eLife.10809.016