title: Cytoplasmic bacteria can be targets for autophagy.
abstract: Autophagy is an important constitutive cellular process involved in size regulation, protein turnover and the removal of malformed or superfluous subcellular components. The process involves the sequestration of cytoplasm and organelles into double-membrane autophagic vacuoles for subsequent breakdown within lysosomes. In this work, we demonstrate that the intracellular pathogen Listeria monocytogenes can also be a target for autophagy. If infected macrophages are treated with chloramphenicol after phagosome lysis, the bacteria are internalized from the cell cytoplasm into autophagic vacuoles. The autophagic vacuoles appear to form by fusion of small cytoplasmic vesicles around the bacteria. These vesicular structures immunolabel with antibodies to protein disulphide isomerase, a marker for the rough ER. Internalization of metabolically arrested cytoplasmic L. monocytogenes represents an autophagic process as the vacuoles have double membranes and the process can be inhibited by the autophagy inhibitors 3-methyladenine and wortmannin. Additionally, the rate of internalization can be accelerated under starvation conditions and the vacuoles fuse with the endocytic pathway. Metabolic inhibition of cytoplasmic bacteria prevents them from adapting to the intracellular niche and reveals a host mechanism utilizing the autophagic pathway as a defence against invading pathogens by providing a route for their removal from the cytoplasm and subsequent delivery to the endocytic pathway for degradation.
authors.affiliation: Ahmanson Advanced Electron Microscopy and Imaging Center, House Ear Institute, and Department of Pathology, Keck School of Medicine, USC, Los Angeles, CA, USA.
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Rich
authors.forename: Chelsea
authors.initials: C
authors.lastname: Burkett
authors.forename: Paul
authors.initials: P
authors.lastname: Webster
mesh_terms: D000818:Animals; D001343:Autophagy; D002460:Cell Line; D002701:Chloramphenicol / Q000494:pharmacology; D003593:Cytoplasm / Q000382:microbiology* / Q000648:ultrastructure; D008089:Listeria monocytogenes / Q000254:growth & development / Q000502:physiology*; D008088:Listeriosis / Q000276:immunology* / Q000382:microbiology; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014617:Vacuoles / Q000382:microbiology
chemical_list: D011500:Protein Synthesis Inhibitors; D002701:Chloramphenicol