title: Intracellular Salmonella induces aggrephagy of host endomembranes in persistent infections.
abstract: Xenophagy has been studied in epithelial cells infected with Salmonella enterica serovar Typhimurium (S. Typhimurium). Distinct autophagy receptors target this pathogen to degradation after interacting with ubiquitin on the surface of cytosolic bacteria, and the phagophore- and autophagosome-associated protein MAP1LC3/LC3. Glycans exposed in damaged phagosomal membranes and diacylglycerol accumulation in the phagosomal membrane also trigger S. Typhimurium xenophagy. How these responses control intraphagosomal and cytosolic bacteria remains poorly understood. Here, we examined S. Typhimurium interaction with autophagy in fibroblasts, in which the pathogen displays limited growth and does not escape into the cytosol. Live-cell imaging microscopy revealed that S. Typhimurium recruits late endosomal or lysosomal compartments that evolve into a membranous aggregate connected to the phagosome. Active dynamics and integrity of the phagosomal membrane are requisite to induce such aggregates. This membranous structure increases over time to become an aggresome that engages autophagy machinery at late infection times (> 6 h postentry). The newly formed autophagosome harbors LC3 and the autophagy receptor SQSTM1/p62 but is devoid of ubiquitin and the receptor CALCOCO2/NDP52. Live-cell imaging showed that this autophagosome captures and digests within the same vacuole the aggresome and some apposed intraphagosomal bacteria. Other phagosomes move away from the aggresome and avoid destruction. Thus, host endomembrane accumulation resulting from activity of intracellular S. Typhimurium stimulates a novel type of aggrephagy that acts independently of ubiquitin and CALCOCO2, and destroys only a few bacteria. Such selective degradation might allow the pathogen to reduce its progeny and, as a consequence, to establish persistent infections.
authors.affiliation: a Laboratory of Intracellular Bacterial Pathogens, Department of Microbial Biotechnology, Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC) , Madrid , Spain.
authors.forename: Noelia
authors.initials: N
authors.lastname: López-Montero
authors.affiliation: a Laboratory of Intracellular Bacterial Pathogens, Department of Microbial Biotechnology, Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC) , Madrid , Spain.
authors.forename: Estel
authors.initials: E
authors.lastname: Ramos-Marquès
authors.affiliation: b Cell Structure Laboratory, Department of Macromolecular Structures, Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC) , Madrid , Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Risco
authors.affiliation: a Laboratory of Intracellular Bacterial Pathogens, Department of Microbial Biotechnology, Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC) , Madrid , Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: García-Del Portillo
mesh_terms: D000818:Animals; D001343:Autophagy; D001426:Bacterial Proteins / Q000378:metabolism; D058947:Bacterial Secretion Systems; D002462:Cell Membrane / Q000473:pathology / Q000648:ultrastructure; D018899:CpG Islands / Q000235:genetics; D005347:Fibroblasts / Q000382:microbiology / Q000473:pathology; D006023:Glycoproteins / Q000378:metabolism; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D042541:Intracellular Space / Q000382:microbiology*; D008247:Lysosomes / Q000378:metabolism / Q000648:ultrastructure; D022101:Microtubule-Organizing Center / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D008954:Models, Biological; D010588:Phagosomes / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D012480:Salmonella Infections / Q000382:microbiology* / Q000473:pathology*; D012486:Salmonella typhimurium / Q000502:physiology*; D012995:Solubility; D025801:Ubiquitin / Q000378:metabolism; D014617:Vacuoles / Q000382:microbiology / Q000648:ultrastructure
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D006023:Glycoproteins; D025801:Ubiquitin
keywords: LGP-membranous aggregate; Salmonella; aggrephagy; fibroblast; persistence; xenophagy