title: The parasitophorous vacuole membrane of Toxoplasma gondii is targeted for disruption by ubiquitin-like conjugation systems of autophagy.
abstract: Autophagy is a lysosomal degradation pathway that is important in cellular homeostasis. Prior work showed a key role for the autophagy related 5 (Atg5) in resistance to Toxoplasma gondii. Here we show that the cassette of autophagy proteins involved in the conjugation of microtubule-associated protein 1 light chain 3 (LC3) to phosphatidylethanolamine, including Atg7, Atg3, and the Atg12-Atg5-Atg16L1 complex play crucial roles in the control of T. gondii in vitro and in vivo. In contrast, pharmacologic modulation of the degradative autophagy pathway or genetic deletion of other essential autophagy genes had no substantial effects. Rather the conjugation system was required for targeting of LC3 and interferon-γ effectors onto the vacuolar membrane of T. gondii and its consequent disruption. These data suggest that the ubiquitin-like conjugation systems that reorganize intracellular membranes during canonical autophagy are necessary for proper targeting of immune effectors to the intracellular vacuole membranes utilized by pathogens.
authors.affiliation: Department of Pathology, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Jayoung
authors.initials: J
authors.lastname: Choi
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Sunmin
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Microbiology, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Scott B
authors.initials: SB
authors.lastname: Biering
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Selleck
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Catherine Y
authors.initials: CY
authors.lastname: Liu
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Genetics, Graduate School of Medicine, WPI Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
authors.forename: Naonobu
authors.initials: N
authors.lastname: Fujita
authors.affiliation: Laboratory of Host Defense, WPI Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Saitoh
authors.affiliation: Laboratory of Host Defense, WPI Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
authors.forename: Shizuo
authors.initials: S
authors.lastname: Akira
authors.affiliation: Department of Genetics, Graduate School of Medicine, WPI Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Yoshimori
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: L David
authors.initials: LD
authors.lastname: Sibley
authors.affiliation: Department of Pathology, University of Chicago, Chicago, IL 60637, USA. Electronic address: shwang@bsd.uchicago.edu.
authors.forename: Seungmin
authors.initials: S
authors.lastname: Hwang
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA. Electronic address: virgin@wustl.edu.
authors.forename: Herbert W
authors.initials: HW
authors.lastname: Virgin
mesh_terms: D000818:Animals; D001343:Autophagy / Q000276:immunology*; D000071192:Autophagy-Related Protein 12; D000071187:Autophagy-Related Protein 5; D000071193:Autophagy-Related Protein 7; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000276:immunology; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011485:Protein Binding / Q000276:immunology; D011506:Proteins / Q000276:immunology; D014122:Toxoplasma / Q000276:immunology*; D014123:Toxoplasmosis / Q000276:immunology* / Q000469:parasitology; D025801:Ubiquitin / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000276:immunology; D014617:Vacuoles / Q000276:immunology / Q000378:metabolism / Q000469:parasitology
chemical_list: C491971:Atg12 protein, mouse; C532820:Atg16l1 protein, mouse; C495773:Atg5 protein, mouse; C502661:Atg7 protein, mouse; D000071192:Autophagy-Related Protein 12; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D010714:Phosphatidylethanolamines; D011506:Proteins; D025801:Ubiquitin; C483858:phosphatidylethanolamine; D007371:Interferon-gamma; D044763:Ubiquitin-Conjugating Enzymes; D000071193:Autophagy-Related Protein 7; C568853:Atg3 protein, mouse