title: Analysis of carbon substrates used by Listeria monocytogenes during growth in J774A.1 macrophages suggests a bipartite intracellular metabolism.
abstract: Intracellular bacterial pathogens (IBPs) are dependent on various nutrients provided by the host cells. Different strategies may therefore be necessary to adapt the intracellular metabolism of IBPs to the host cells. The specific carbon sources, the catabolic pathways participating in their degradation, and the biosynthetic performances of IBPs are still poorly understood. In this report, we have exploited the technique of (13)C-isotopologue profiling to further study the carbon metabolism of Listeria monocytogenes by using the EGDe wild-type strain and mutants (defective in the uptake and/or catabolism of various carbon compounds) replicating in J774A.1 macrophages. For this goal, the infected macrophages were cultivated in the presence of [1,2-(13)C2]glucose, [U-(13)C3]glycerol, [U-(13)C3]pyruvate, [U-(13)C3]lactate, or a mix of [U-(13)C]amino acids. GC/MS-based isotopologue profiling showed efficient utilization of amino acids, glucose 6-phosphate, glycerol, and (at a low extent) also of lactate but not of pyruvate by the IBPs. Most amino acids imported from the host cells were directly used for bacterial protein biosynthesis and hardly catabolized. However, Asp was de novo synthesized by the IBPs and not imported from the host cell. As expected, glycerol was catabolized via the ATP-generating lower part of the glycolytic pathway, but apparently not used for gluconeogenesis. The intermediates generated from glucose 6-phosphate in the upper part of the glycolytic pathway and the pentose phosphate shunt likely serve primarily for anabolic purposes (probably for the biosynthesis of cell wall components and nucleotides). This bipartite bacterial metabolism which involves at least two major carbon substrates-glycerol mainly for energy supply and glucose 6-phosphate mainly for indispensible anabolic performances-may put less nutritional stress on the infected host cells, thereby extending the lifespan of the host cells to the benefit of the IBPs.
authors.affiliation: Lehrstuhl für Biochemie, Technische Universität München Garching, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Grubmüller
authors.affiliation: Abteilung Mikrobiologie, Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Technische Universität München Freising, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Schauer
authors.affiliation: Department for Bacteriology, Max von Pettenkofer Institute, Ludwig-Maximilians-Universität München, Germany.
authors.forename: Werner
authors.initials: W
authors.lastname: Goebel
authors.affiliation: Abteilung Mikrobiologie, Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Technische Universität München Freising, Germany.
authors.forename: Thilo M
authors.initials: TM
authors.lastname: Fuchs
authors.affiliation: Lehrstuhl für Biochemie, Technische Universität München Garching, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Eisenreich
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002244:Carbon / Q000378:metabolism*; D002384:Catalysis; D002460:Cell Line; D005943:Gluconeogenesis; D019298:Glucose-6-Phosphate / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D054884:Host-Pathogen Interactions; D019344:Lactic Acid / Q000378:metabolism; D008089:Listeria monocytogenes / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology / Q000382:microbiology*; D051379:Mice; D009154:Mutation; D019289:Pyruvic Acid / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D019344:Lactic Acid; D019298:Glucose-6-Phosphate; D002244:Carbon; D019289:Pyruvic Acid; D005990:Glycerol
keywords: Listeria monocytogenes; bacterial metabolism; bacterial pathogensis; intracellular bacteria; isotopic tracers; isotopologue profiling