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Proximity-labeling reveals novel host and parasite proteins at the Toxoplasma parasitophorous vacuole membrane

ABSTRACT: Cygan AM, Jean Beltran PM, Branon TC, Ting AY, Carr SA, Boothroyd JC. 2021. Toxoplasma gondii is a ubiquitous, intracellular parasite that envelopes its parasitophorous vacuole with a protein-laden membrane (PVM). The PVM is critical for interactions with the infected host cell such as nutrient transport and immune defense. Only a few parasite and host proteins have so far been identified on the host-cytosolic side of the PVM. We report here the use of human foreskin fibroblasts expressing the proximity-labeling enzyme miniTurbo, fused to a domain that targets it to this face of the PVM, in combination with quantitative proteomics to specifically identify proteins present at this crucial interface. Out of numerous host and parasite proteins with candidate PVM localization, we validate three novel parasite proteins (TGGT1_269950, TGGT1_215360, and TGGT1_217530) and four new host proteins (PDCD6IP/ALIX, PDCD6, CC2D1A, and MOSPD2) as PVM-localized in infected human cells through immunofluorescent microscopy. These results significantly expand our knowledge of proteins present at the PVM and, given that three of the validated host proteins are components of the ESCRT machinery, they further suggest that novel biology is operating at this crucial host-pathogen interface.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Toxoplasma Gondii (ncbitaxon:5811) Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Steven A. Carr

PROVIDER: MSV000086833 | MassIVE | Wed Feb 10 06:59:00 GMT 2021

REPOSITORIES: MassIVE

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