@article{471f57c52fed42b9bbc7081204a60da6,
title = "The parasite Toxoplasma sequesters diverse Rab host vesicles within an intravacuolar network",
abstract = "Many intracellular pathogens subvert host membrane trafficking pathways to promote their replication. Toxoplasma multiplies in a membrane-bound parasitophorous vacuole (PV) that interacts with mammalian host organelles and intercepts Golgi Rab vesicles to acquire sphingolipids. The mechanisms of host vesicle internalization and processing within the PV remain undefined. We demonstrate that Toxoplasma sequesters a broad range of Rab vesicles into the PV. Correlative light and electron microscopy analysis of infected cells illustrates that intravacuolar Rab1A vesicles are surrounded by the PV membrane, suggesting a phagocytic-like process for vesicle engulfment. Rab11A vesicles concentrate to an intravacuolar network (IVN), but this is reduced in Δgra2 and Δgra2Δgra6 parasites, suggesting that tubules stabilized by the TgGRA2 and TgGRA6 proteins secreted by the parasite within the PV contribute to host vesicle sequestration. Overexpression of a phospholipase TgLCAT, which is localized to the IVN, results in a decrease in the number of intravacuolar GFP-Rab11A vesicles, suggesting that TgLCAT controls lipolytic degradation of Rab vesicles for cargo release.",
author = "Romano, {Julia D.} and Nolan, {Sabrina J.} and Corey Porter and Karen Ehrenman and Hartman, {Eric J.} and Hsia, {Ru ching} and Isabelle Coppens",
note = "Funding Information: This work used an EM sample preparation instrument that was purchased with funding from a National Institutes of Health Shared Instrumentation Grant (1S10RR26870-1) awarded to University of Maryland, Baltimore. The mouse monoclonal anti–LIMP/LAMP3/ CD63 (H5C6) antibody developed by Thomas August and James E.K. Hildreth was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the Eunice Kennedy Shriver National Institute of Child Health and Human Development and maintained by the Department of Biology, The University of Iowa, Iowa City, IA. This study was supported by the National Institutes of Health (grant AI060767 to I. Coppens). The authors declare no competing financial interests. Funding Information: We thank the members of the Coppens' laboratory for helpful discussion during the course of this work. We are grateful to the generous providers of plasmids and antibodies used in this study. We also thank the excellent technical staff of the Electron Microscopy Core Facility at Yale School of Medicine, the Johns Hopkins University School of Medicine Microscopy Facility, and John Strong at the Electron Microscopy Core Imaging Facility of the University of Maryland. This work used an EM sample preparation instrument that was purchased with funding from a National Institutes of Health Shared Instrumentation Grant (1S10RR26870-1) awarded to University of Maryland, Baltimore. The mouse monoclonal anti-LIMP/LAMP3/ CD63 (H5C6) antibody developed by Thomas August and James E.K. Hildreth was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the Eunice Kennedy Shriver National Institute of Child Health and Human Development and maintained by the Department of Biology, The University of Iowa, Iowa City, IA. This study was supported by the National Institutes of Health (grant AI060767 to I. Coppens) Publisher Copyright: {\textcopyright} 2017 Romano et al.",
year = "2017",
doi = "10.1083/jcb.201701108",
language = "English (US)",
volume = "216",
pages = "4235--4254",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "12",
}