title: MTOR-Driven Metabolic Reprogramming Regulates Legionella pneumophila Intracellular Niche Homeostasis.
abstract: Vacuolar bacterial pathogens are sheltered within unique membrane-bound organelles that expand over time to support bacterial replication. These compartments sequester bacterial molecules away from host cytosolic immunosurveillance pathways that induce antimicrobial responses. The mechanisms by which the human pulmonary pathogen Legionella pneumophila maintains niche homeostasis are poorly understood. We uncovered that the Legionella-containing vacuole (LCV) required a sustained supply of host lipids during expansion. Lipids shortage resulted in LCV rupture and initiation of a host cell death response, whereas excess of host lipids increased LCVs size and housing capacity. We found that lipids uptake from serum and de novo lipogenesis are distinct redundant supply mechanisms for membrane biogenesis in Legionella-infected macrophages. During infection, the metabolic checkpoint kinase Mechanistic Target of Rapamycin (MTOR) controlled lipogenesis through the Serum Response Element Binding Protein 1 and 2 (SREBP1/2) transcription factors. In Legionella-infected macrophages a host-driven response that required the Toll-like receptors (TLRs) adaptor protein Myeloid differentiation primary response gene 88 (Myd88) dampened MTOR signaling which in turn destabilized LCVs under serum starvation. Inactivation of the host MTOR-suppression pathway revealed that L. pneumophila sustained MTOR signaling throughout its intracellular infection cycle by a process that required the upstream regulator Phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) and one or more Dot/Icm effector proteins. Legionella-sustained MTOR signaling facilitated LCV expansion and inhibition of the PI3K-MTOR-SREPB1/2 axis through pharmacological or genetic interference or by activation of the host MTOR-suppression response destabilized expanding LCVs, which in turn triggered cell death of infected macrophages. Our work identified a host metabolic requirement for LCV homeostasis and demonstrated that L. pneumophila has evolved to manipulate MTOR-dependent lipogenesis for optimal intracellular replication.
authors.affiliation: Department of Medicine, Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center - Shreveport, Shreveport, Louisiana, United States of America.
authors.forename: Camille F
authors.initials: CF
authors.lastname: Abshire
authors.affiliation: Department of Medicine, Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center - Shreveport, Shreveport, Louisiana, United States of America.
authors.forename: Ana-Maria
authors.identifier: http://orcid.org/0000-0001-9841-6802
authors.initials: AM
authors.lastname: Dragoi
authors.affiliation: Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, Connecticut, United States of America.
authors.forename: Craig R
authors.initials: CR
authors.lastname: Roy
authors.affiliation: Department of Microbiology and Immunology, Louisiana State University Health Sciences Center - Shreveport, Shreveport, Louisiana, United States of America.
authors.forename: Stanimir S
authors.identifier: http://orcid.org/0000-0002-3294-7595
authors.initials: SS
authors.lastname: Ivanov
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005455:Fluorescent Antibody Technique; D006706:Homeostasis / Q000502:physiology; D054884:Host-Pathogen Interactions / Q000502:physiology*; D021621:Imaging, Three-Dimensional; D015151:Immunoblotting; D016952:Legionella pneumophila / Q000378:metabolism*; D007877:Legionnaires' Disease / Q000378:metabolism* / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008853:Microscopy / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism
chemical_list: D058570:TOR Serine-Threonine Kinases