@article{fb3873bd16124723a10bc88004d1fa66,
title = "Toxoplasma gondii exploits the host ESCRT machinery for parasite uptake of host cytosolic proteins",
abstract = "Toxoplasma gondii is a master manipulator capable of effectively siphoning the resources from the host cell for its intracellular subsistence. However, the molecular underpinnings of how the parasite gains resources from its host remain largely unknown. Residing within a non-fusogenic parasitophorous vacuole (PV), the parasite must acquire resources across the limiting membrane of its replicative niche, which is decorated with parasite proteins including those secreted from dense granules. We discovered a role for the Endosomal Sorting Complex Required for Transport (ESCRT) machinery in host cytosolic protein uptake by T. gondii by disrupting host ESCRT function. We identified the transmembrane dense granule protein TgGRA14, which contains motifs homologous to the late domain motifs of HIV-1 Gag, as a candidate for the recruitment of the host ESCRT machinery to the PV membrane. Using an HIV-1 virus-like particle (VLP) release assay, we found that the motif-containing portion of TgGRA14 is sufficient to substitute for HIV-1 Gag late domain to mediate ESCRT-dependent VLP budding. We also show that TgGRA14 is proximal to and interacts with host ESCRT components and other dense granule proteins during infection. Furthermore, analysis of TgGRA14-deficient parasites revealed a marked reduction in ingestion of a host cytosolic protein compared to WT parasites. Thus, we propose a model in which T. gondii recruits the host ESCRT machinery to the PV where it can interact with TgGRA14 for the internalization of host cytosolic proteins across the PV membrane (PVM). These findings provide new insight into how T. gondii accesses contents of the host cytosol by exploiting a key pathway for vesicular budding and membrane scission.",
author = "Yolanda Rivera-Cuevas and Joshua Mayoral and {Di Cristina}, Manlio and Lawrence, {Anna Lisa E.} and Olafsson, {Einar B.} and Patel, {Romir K.} and Dishari Thornhill and Waldman, {Benjamin S.} and Akira Ono and Sexton, {Jonathan Z.} and Sebastian Lourido and Weiss, {Louis M.} and Carruthers, {Vern B.}",
note = "Funding Information: We gratefully acknowledge the funding support from U.S. National Institutes of Health grants T32AI007528 (Y.R.C.), F31AI152297 (Y.R. C), F31AI36401 (J.M.), T32GM007288 (J.M.), the University of Perugia Fondo Ricerca Di Base 2019 program of the Department of Chemistry, Biology, and Biotechnology (M.D.C.), the U.S. National Institutes of Health grant T32AI007414 (E.B.O.), University of Michigan Life Sciences Fellowship Program (E.B.O.), U.S. National Institutes of Health grants R37AI071727 (A.O.), R01AI158501 (S.L.), and R01DK120623 (J.Z.S.), the University of Michigan Institute for Clinical and Health Research grant UL1TR002240 (J.Z.S.), U.S. National Institutes of Health grant P30DK034933 (J.Z.S.), the Edward Mallinckrodt, Jr. Foundation (S.L), and U.S. National Institutes of Health grants R01AI34753 (L.M.W.) and R01AI120607 (V.B.C). We thank Dr. Peter Bradley for providing us with the pGRA14-HA plasmid and the RΔgra14 and R:GRA14OE parasite strains and Dr. Schuyler B. van Engelenburg for sharing the endogenously tagged GFP-TSG101 HeLa cells with us. We also thank Drs. Yoshifumi Nishikawa, Gary Ward, Furio Spano, L. David Sibley, Jeroen Saeij, Peter Bradley, and Dominique Soldati-Favre for providing antibodies along with Drs. Phyllis Hanson and Isabelle Coppens for their input for this project. Publisher Copyright: {\textcopyright} 2021 Rivera-Cuevas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",
year = "2021",
month = dec,
doi = "10.1371/journal.ppat.1010138",
language = "English (US)",
volume = "17",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "12",
}