title: Toxoplasma aldolase is required for metabolism but dispensable for host-cell invasion.
abstract: Gliding motility and host-cell invasion by apicomplexan parasites depend on cell-surface adhesins that are translocated via an actin-myosin motor beneath the membrane. The current model posits that fructose-1,6-bisphosphate aldolase (ALD) provides a critical link between the cytoplasmic tails of transmembrane adhesins and the actin-myosin motor. Here we tested this model using the Toxoplasma gondii apical membrane protein 1 (TgAMA1), which binds to aldolase in vitro. TgAMA1 cytoplasmic tail mutations that disrupt ALD binding in vitro showed no correlation with host-cell invasion, indicating this interaction is not essential. Furthermore, ALD-depleted parasites were impaired when grown in glucose, yet they showed normal gliding and invasion in glucose-free medium. Depletion of ALD in the presence of glucose led to accumulation of fructose-1,6-bisphosphate, which has been associated with toxicity in other systems. Finally, TgALD knockout parasites and an ALD mutant that specifically disrupts adhesin binding in vitro also supported normal invasion when cultured in glucose-free medium. Taken together, these results suggest that ALD is primarily important for energy metabolism rather than interacting with microneme adhesins, challenging the current model for apicomplexan motility and invasion.
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Bang
authors.initials: B
authors.lastname: Shen
authors.forename: L David
authors.initials: LD
authors.lastname: Sibley
mesh_terms: D015153:Blotting, Western; D002853:Chromatography, Liquid; D018076:DNA, Complementary / Q000235:genetics; D004734:Energy Metabolism / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005634:Fructose-Bisphosphate Aldolase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D055786:Gene Knockout Techniques; D005947:Glucose; D006790:Host-Parasite Interactions / Q000502:physiology*; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D010957:Plasmids / Q000235:genetics; D053719:Tandem Mass Spectrometry; D014122:Toxoplasma / Q000201:enzymology* / Q000502:physiology
chemical_list: D018076:DNA, Complementary; D005634:Fructose-Bisphosphate Aldolase; D005947:Glucose
keywords: glycolysis; micronemal adhesin; motor complex