Glutamate Utilization Couples Oxidative Stress Defense and the Tricarboxylic Acid Cycle in Phagosomal Escape

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Intracellular bacterial pathogens have developed a variety of strategies to avoid degradation by the host innate immune defense mechanisms triggered upon phagocytocis. Upon infection of mammalian host cells, the intracellular pathogen Francisella replicates exclusively in the cytosolic compartment. Hence, its ability to escape rapidly from the phagosomal compartment is critical for its pathogenicity. Here, we show for the first time that a glutamate transporter of Francisella (here designated GadC) is critical for oxidative stress defense in the phagosome, thus impairing intra-macrophage multiplication and virulence in the mouse model. The gadC mutant failed to efficiently neutralize the production of reactive oxygen species. Remarkably, virulence of the gadC mutant was partially restored in mice defective in NADPH oxidase activity. The data presented highlight links between glutamate uptake, oxidative stress defense, the tricarboxylic acid cycle and phagosomal escape. This is the first report establishing the role of an amino acid transporter in the early stage of the Francisella intracellular lifecycle.

Vyšlo v časopise: Glutamate Utilization Couples Oxidative Stress Defense and the Tricarboxylic Acid Cycle in Phagosomal Escape. PLoS Pathog 10(1): e32767. doi:10.1371/journal.ppat.1003893
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003893

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Zdroje

1. Sjostedt A, editor (2011) Francisella tularensis and tularemia: Fontiers Media SA.

2. OystonPC, SjostedtA, TitballRW (2004) Tularaemia: bioterrorism defence renews interest in Francisella tularensis. Nat Rev Microbiol 2 : 967–978.

3. KeimP, JohanssonA, WagnerDM (2007) Molecular epidemiolgy, evolution, and ecology of Francisella. Ann N Y Acad Sci 1105 : 30–66.

4. CelliJ, ZahrtTC (2013) Mechanisms of Francisella tularensis intracellular pathogenesis. Cold Spring Harb Perspect Med 3: a010314.

5. McCaffreyRL, SchwartzJT, LindemannSR, MorelandJG, BuchanBW, et al. (2010) Multiple mechanisms of NADPH oxidase inhibition by type A and type B Francisella tularensis. J Leukoc Biol 88 : 791–805.

6. MeibomKL, CharbitA (2010) The unraveling panoply of Francisella tularensis virulence attributes. Curr Opin Microbiol 13 : 11–17.

7. NapierBA, MeyerL, BinaJE, MillerMA, SjostedtA, et al. (2012) Link between intraphagosomal biotin and rapid phagosomal escape in Francisella. Proc Natl Acad Sci USA 109 : 18084–18089.

8. MelilloAA, BakshiCS, MelendezJA (2010) Francisella tularensis antioxidants harness reactive oxygen species to restrict macrophage signaling and cytokine production. J Biol Chem 285 : 27553–27560.

9. LindgrenH, ShenH, ZingmarkC, GolovliovI, ConlanW, et al. (2007) Resistance of Francisella strains against reactive nitrogen and oxygen species with special reference to the role of KatG. Infect Immun 75 : 1303–1309.

10. MelilloAA, MahawarM, SellatiTJ, MalikM, MetzgerDW, et al. (2009) Identification of Francisella tularensis live vaccine strain CuZn superoxide dismutase as critical for resistance to extracellularly generated reactive oxygen species. J Bacteriol 191 : 6447–6456.

11. BakshiCS, MalikM, ReganK, MelendezJA, MetzgerDW, et al. (2006) Superoxide dismutase B gene (sodB)-deficient mutants of Francisella tularensis demonstrate hypersensitivity to oxidative stress and attenuated virulence. J Bacteriol 188 : 6443–6448.

12. DieppedaleJ, GesbertG, RamondE, ChhuonC, DubailI, et al. (2013) Possible links between stress defense and the tricarboxylic acid cycle in Francisella pathogenesis. Mol Cell Proteomics 12 : 2278–2292.

13. ClemensDL, LeeBY, HorwitzMA (2004) Virulent and avirulent strains of Francisella tularensis prevent acidification and maturation of their phagosomes and escape into the cytoplasm in human macrophages. Infect Immun 72 : 3204–3217.

14. ClemensDL, LeeBY, HorwitzMA (2009) Francisella tularensis phagosomal escape does not require acidification of the phagosome. Infect Immun 77 : 1757–1773.

15. SanticM, AsareR, SkrobonjaI, JonesS, Abu KwaikY (2008) Acquisition of the vacuolar ATPase proton pump and phagosome acidification are essential for escape of Francisella tularensis into the macrophage cytosol. Infect Immun 76 : 2671–2677.

16. De BiaseD, PennacchiettiE (2012) Glutamate decarboxylase-dependent acid resistance in orally acquired bacteria: function, distribution and biomedical implications of the gadBC operon. Mol Microbiol 86 (4) 770–86.

17. FosterJW (2004) Escherichia coli acid resistance: tales of an amateur acidophile. Nat Rev Microbiol 2 : 898–907.

18. KaratzasKA, BrennanO, HeavinS, MorrisseyJ, O'ByrneCP (2010) Intracellular accumulation of high levels of gamma-aminobutyrate by Listeria monocytogenes 10403S in response to low pH: uncoupling of gamma-aminobutyrate synthesis from efflux in a chemically defined medium. Appl Environ Microbiol 76 : 3529–3537.

19. MeibomKL, CharbitA (2010) Francisella tularensis metabolism and its relation to virulence. Front Microbiol 1 : 140.

20. GesbertG, RamondE, RigardM, FrapyE, DupuisM, et al. (2013) Asparagine assimilation is critical for intracellular replication and dissemination of Francisella. Cell Microbiol [epub ahead of print].

21. MaierTM, CaseyMS, BeckerRH, DorseyCW, GlassEM, et al. (2007) Identification of Francisella tularensis Himar1-based transposon mutants defective for replication in macrophages. Infect Immun 75 : 5376–5389.

22. WeissDS, BrotckeA, HenryT, MargolisJJ, ChanK, et al. (2007) In vivo negative selection screen identifies genes required for Francisella virulence. Proc Natl Acad Sci USA 104 : 6037–6042.

23. PengK, MonackDM (2010) Indoleamine 2,3-dioxygenase 1 is a lung-specific innate immune defense mechanism that inhibits growth of Francisella tularensis tryptophan auxotrophs. Infect Immun 78 : 2723–2733.

24. KraemerPS, MitchellA, PelletierMR, GallagherLA, WasnickM, et al. (2009) Genome-wide screen in Francisella novicida for genes required for pulmonary and systemic infection in mice. Infect Immun 77 : 232–244.

25. MaD, LuP, YanC, FanC, YinP, et al. (2012) Structure and mechanism of a glutamate-GABA antiporter. Nature 483 : 632–636.

26. LaurianoCM, BarkerJR, NanoFE, ArulanandamBP, KloseKE (2003) Allelic exchange in Francisella tularensis using PCR products. FEMS Microbiol Lett 229 : 195–202.

27. ChamberlainRE (1965) Evaluation of Live Tularemia Vaccine Prepared in a Chemically Defined Medium. Appl Microbiol 13 : 232–235.

28. BarelM, MeibomK, CharbitA (2010) Nucleolin, a shuttle protein promoting infection of human monocytes by Francisella tularensis. PLoS One 5: e14193.

29. ChongA, WehrlyT, ChildR, HansenB, HwangS, et al. (2012) Cytosolic clearance of replication-deficient mutants reveals Francisella tularensis interactions with the autophagic pathway. Autophagy 8 : 1342–1356.

30. Fernandes-AlnemriT, YuJW, JulianaC, SolorzanoL, KangS, et al. (2010) The AIM2 inflammasome is critical for innate immunity to Francisella tularensis. Nat Immunol 11 : 385–393.

31. JonesJW, BrozP, MonackDM (2011) Innate immune recognition of Francisella tularensis: activation of type-I interferons and the inflammasome. Front Microbiol 2 : 16.

32. RathinamVA, JiangZ, WaggonerSN, SharmaS, ColeLE, et al. (2010) The AIM2 inflammasome is essential for host defense against cytosolic bacteria and DNA viruses. Nat Immunol 11 : 395–402.

33. PieriniR, JurujC, PerretM, JonesCL, MangeotP, et al. (2012) AIM2/ASC triggers caspase-8-dependent apoptosis in Francisella-infected caspase-1-deficient macrophages. Cell Death Differ 19 : 1709–1721.

34. JackDL, PaulsenIT, SaierMH (2000) The amino acid/polyamine/organocation (APC) superfamily of transporters specific for amino acids, polyamines and organocations. Microbiology 146 (Pt 8) 1797–1814.

35. TakanashiT, OguraY, TaguchiH, HashizoeM, HondaY (1997) Fluorophotometric quantitation of oxidative stress in the retina in vivo. Invest Ophthalmol Vis Sci 38 : 2721–2728.

36. KwaikYA, BumannD (2013) Microbial Quest for Food in vivo: “Nutritional virulence” as an emerging paradigm. Cell Microbiol 15 : 882–890.

37. MohapatraNP, SoniS, ReillyTJ, LiuJ, KloseKE, et al. (2008) Combined deletion of four Francisella novicida acid phosphatases attenuates virulence and macrophage vacuolar escape. Infect Immun 76 : 3690–3699.

38. MohapatraNP, SoniS, RajaramMV, StrandbergKL, GunnJS (2013) Type A Francisella tularensis Acid Phosphatases Contribute to Pathogenesis. PLoS One 8: e56834.

39. NewsholmeP, ProcopioJ, LimaMM, Pithon-CuriTC, CuriR (2003) Glutamine and glutamate–their central role in cell metabolism and function. Cell Biochem Funct 21 : 1–9.

40. FeehilyC, KaratzasKA (2012) Role of glutamate metabolism in bacterial responses towards acid and other stresses. J Appl Microbiol 114 : 11–24.

41. CaoJ, BarbosaJM, SinghNK, LocyRD (2013) GABA shunt mediates thermotolerance in Saccharomyces cerevisiae by reducing reactive oxygen production. Yeast 30 : 129–144.

42. KaratzasKA, SuurL, O'ByrneCP (2012) Characterization of the intracellular glutamate decarboxylase system: analysis of its function, transcription, and role in the acid resistance of various strains of Listeria monocytogenes. Appl Environ Microbiol 78 : 3571–3579.

43. AsareR, AkimanaC, JonesS, Abu KwaikY (2010) Molecular bases of proliferation of Francisella tularensis in arthropod vectors. Environ Microbiol 12 : 2587–2612.

44. MaillouxRJ, BeriaultR, LemireJ, SinghR, ChenierDR, et al. (2007) The tricarboxylic acid cycle, an ancient metabolic network with a novel twist. PLoS One 2: e690.

45. EohH, RheeKY (2013) Multifunctional essentiality of succinate metabolism in adaptation to hypoxia in Mycobacterium tuberculosis. Proc Natl Acad Sci U S A 110 : 6554–6559.

46. RichardsonAR, PayneEC, YoungerN, KarlinseyJE, ThomasVC, et al. (2011) Multiple targets of nitric oxide in the tricarboxylic acid cycle of Salmonella enterica serovar typhimurium. Cell Host Microbe 10 : 33–43.

47. ThompsonLJ, MerrellDS, NeilanBA, MitchellH, LeeA, et al. (2003) Gene expression profiling of Helicobacter pylori reveals a growth-phase-dependent switch in virulence gene expression. Infect Immun 71 : 2643–2655.

48. AlkhuderK, MeibomKL, DubailI, DupuisM, CharbitA (2009) Glutathione provides a source of cysteine essential for intracellular multiplication of Francisella tularensis. PLoS Pathog 5: e1000284.

49. Castanie-CornetMP, PenfoundTA, SmithD, ElliottJF, FosterJW (1999) Control of acid resistance in Escherichia coli. J Bacteriol 181 : 3525–3535.

50. RebrinI, KamzalovS, SohalRS (2003) Effects of age and caloric restriction on glutathione redox state in mice. Free Radic Biol Med 35 : 626–635.

51. BrotckeA, WeissDS, KimCC, ChainP, MalfattiS, et al. (2006) Identification of MglA-regulated genes reveals novel virulence factors in Francisella tularensis. Infect Immun 74 : 6642–6655.

52. CowingDW, BardwellJC, CraigEA, WoolfordC, HendrixRW, et al. (1985) Consensus sequence for Escherichia coli heat shock gene promoters. Proc Natl Acad Sci U S A 82 : 2679–2683.