title: Coxiella burnetii type IV secretion-dependent recruitment of macrophage autophagosomes.
abstract: Coxiella burnetii is an intracellular Gram-negative bacterium that causes human Q fever, a flu-like disease that can progress to chronic, life-threatening endocarditis. In humans, C. burnetii infects alveolar macrophages and promotes phagosomal fusion with autophagosomes and lysosomes, establishing a unique parasitophorous vacuole (PV) in which to replicate. The pathogen uses a Dot/Icm type IV secretion system (T4SS) to deliver effector proteins to the host cytoplasm, where they alter cellular processes to benefit the pathogen. The T4SS is required for PV expansion and prevention of apoptosis, but little else is known about the role of the system during intracellular growth. Recent reports suggest that C. burnetii actively recruits autophagosomes to the PV to deliver nutrients to the pathogen and provide membrane for the expanding vacuole. In this study, we examined the role of the T4SS in mediating PV interactions with autophagosomes. We found that the autophagy-related proteins LC3 and p62 localized to wild-type PV but not to T4SS mutant organism-containing phagosomes in human macrophage-like cells, primary human alveolar macrophages, and Chinese hamster ovary cells. However, while lipidated LC3 levels were elevated regardless of T4SS activity, no p62 turnover was observed during C. burnetii growth in macrophages, suggesting that the pathogen recruits preformed autophagosomes. When the T4SS was activated 24 h after infection, autophagosome recruitment ensued, indicating that autophagosome interactions are dispensable for initial PV maturation to a phagolysosome-like compartment but are involved in vacuole expansion. Together, these results demonstrate that C. burnetii actively directs PV-autophagosome interactions by using the Dot/Icm T4SS.
authors.affiliation: Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA.
authors.forename: Caylin G
authors.initials: CG
authors.lastname: Winchell
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Graham
authors.forename: Richard C
authors.initials: RC
authors.lastname: Kurten
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Voth
mesh_terms: D001343:Autophagy / Q000502:physiology*; D058947:Bacterial Secretion Systems / Q000502:physiology*; D016997:Coxiella burnetii / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology*; D010588:Phagosomes / Q000378:metabolism / Q000382:microbiology*; D014617:Vacuoles / Q000378:metabolism / Q000382:microbiology
chemical_list: D058947:Bacterial Secretion Systems