title: Functional dissection of Toxoplasma gondii perforin-like protein 1 reveals a dual domain mode of membrane binding for cytolysis and parasite egress.
abstract: The recently discovered role of a perforin-like protein (PLP1) for rapid host cell egress by the protozoan parasite Toxoplasma gondii expanded the functional diversity of pore-forming proteins. Whereas PLP1 was found to be necessary for rapid egress and pathogenesis, the sufficiency for and mechanism of membrane attack were yet unknown. Here we further dissected the PLP1 knock-out phenotype, the mechanism of PLP1 pore formation, and the role of each domain by genetic complementation. We found that PLP1 is sufficient for membrane disruption and has a conserved mechanism of pore formation through target membrane binding and oligomerization to form large, multimeric membrane-embedded complexes. The highly conserved, central MACPF domain and the β-sheet-rich C-terminal domain were required for activity. Loss of the unique N-terminal extension reduced lytic activity and led to a delay in rapid egress, but did not significantly decrease virulence, suggesting that small amounts of lytic activity are sufficient for pathogenesis. We found that both N- and C-terminal domains have membrane binding activity, with the C-terminal domain being critical for function. This dual mode of membrane association may promote PLP1 activity and parasite egress in the diverse cell types in which this parasite replicates.
authors: Cell and Molecular Biology Program, University of Michigan, Ann Arbor, Michigan 48109-5630; Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 48109-5630.
authors: Marijo S
authors: MS
authors: Roiko
authors: Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 48109-5630. Electronic address: vcarruth@umich.edu.
authors: Vern B
authors: VB
authors: Carruthers
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism / Q000469:parasitology; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D004912:Erythrocytes / Q000469:parasitology; D005260:Female; D055786:Gene Knockout Techniques; D006461:Hemolysis; D006790:Host-Parasite Interactions; D006801:Humans; D051379:Mice; D054353:Perforin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D014122:Toxoplasma / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014123:Toxoplasmosis / Q000469:parasitology
chemical_list: D015800:Protozoan Proteins; D054353:Perforin
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