Detection of Host-Derived Sphingosine by Is Important for Survival in the Murine Lung

English version České info

Pseudomonas aeruginosa is a common environmental bacterium that is also a significant opportunistic pathogen, particularly of the human lung. We must understand how P. aeruginosa responds to the lung environment in order to identify the regulatory changes that bacteria use to establish and maintain infections. The P. aeruginosa response to pulmonary surfactant was used as a model to identify transcripts likely induced during lung infection. The most highly induced transcript in pulmonary surfactant, PA5325 (sphA), is regulated by an AraC-family transcription factor, PA5324 (SphR). We found that sphA was specifically induced by sphingosine in an SphR-dependent manner, and also via metabolism of sphingomyelin, ceramide, or sphingoshine-1-phosphate to sphingosine. These sphingolipids not only play a structural role in lipid membranes, but some are also intracellular and intercellular signaling molecules important in normal eukaryotic cell functions as well as orchestrating immune responses. The members of the SphR transcriptome were identified by microarray analyses, and DNA binding assays showed specific interaction of these promoters with SphR, which enabled us to determine the consensus SphR binding site. SphR binding to DNA was modified by sphingosine and we used labeled sphingosine to demonstrate direct binding of sphingosine by SphR. Deletion of sphR resulted in reduced bacterial survival during mouse lung infection. In vitro experiments show that deletion of sphR increases sensitivity to the antimicrobial effects of sphingosine which could, in part, explain the in vivo phenotype. This is the first identification of a sphingosine-responsive transcription factor in bacteria. We predict that SphR transcriptional regulation may be important in response to many sites of infection in eukaryotes and the presence of homologous transcription factors in other pathogens suggests that sphingosine detection is not limited to P. aeruginosa.

Vyšlo v časopise: Detection of Host-Derived Sphingosine by Is Important for Survival in the Murine Lung. PLoS Pathog 10(1): e32767. doi:10.1371/journal.ppat.1003889
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003889

Souhrn

Zdroje

1. LiebermanD, LiebermanD (2003) Pseudomonal infections in patients with COPD: epidemiology and management. Am J Respir Med 2 : 459–468.

2. ChastreJ, FagonJY (2002) Ventilator-associated pneumonia. Am J Respir Crit Care Med 165 : 867–903.

3. Crouch BrewerS, WunderinkRG, JonesCB, LeeperKVJr (1996) Ventilator-associated pneumonia due to Pseudomonas aeruginosa. Chest 109 : 1019–1029.

4. BurnsJL, EmersonJ, StappJR, YimDL, KrzewinskiJ, et al. (1998) Microbiology of sputum from patients at cystic fibrosis centers in the United States. Clin Infect Dis 27 : 158–163.

5. BriesacherBA, QuittnerAL, FouayziH, ZhangJ, SwensenA (2011) Nationwide trends in the medical care costs of privately insured patients with cystic fibrosis (CF), 2001–2007. Pediatr Pulmonol 46 : 770–776.

6. BurnsJL, GibsonRL, McNamaraS, YimD, EmersonJ, et al. (2001) Longitudinal assessment of Pseudomonas aeruginosa in young children with cystic fibrosis. J Infect Dis 183 : 444–452.

7. RajanS, SaimanL (2002) Pulmonary infections in patients with cystic fibrosis. Semin Respir Infect 17 : 47–56.

8. Butorac-PetanjekB, ParnhamMJ, Popovic-GrleS (2010) Antibiotic therapy for exacerbations of chronic obstructive pulmonary disease (COPD). J Chemother 22 : 291–297.

9. KlockgetherJ, MunderA, NeugebauerJ, DavenportCF, StankeF, et al. (2010) Genome diversity of Pseudomonas aeruginosa PAO1 laboratory strains. J Bacteriol 192 : 1113–1121.

10. FischbachMA, WalshCT (2009) Antibiotics for emerging pathogens. Science 325 : 1089–1093.

11. FriskA, SchurrJR, WangG, BertucciDC, MarreroL, et al. (2004) Transcriptome analysis of Pseudomonas aeruginosa after interaction with human airway epithelial cells. Infect Immun 72 : 5433–5438.

12. PalmerKL, MashburnLM, SinghPK, WhiteleyM (2005) Cystic fibrosis sputum supports growth and cues key aspects of Pseudomonas aeruginosa physiology. J Bacteriol 187 : 5267–5277.

13. ChroneosZC, Sever-ChroneosZ, ShepherdVL (2010) Pulmonary surfactant: an immunological perspective. Cell Physiol Biochem 25 : 13–26.

14. GlasserJR, MallampalliRK (2012) Surfactant and its role in the pathobiology of pulmonary infection. Microbes Infect 14 : 17–25.

15. CaminitiSP, YoungSL (1991) The pulmonary surfactant system. Hosp Pract (Off Ed) 26 : 87–90, 94–100.

16. HannunYA, ObeidLM (2008) Principles of bioactive lipid signalling: lessons from sphingolipids. Nat Rev Mol Cell Biol 9 : 139–150.

17. SpiegelS, MilstienS (2011) The outs and the ins of sphingosine-1-phosphate in immunity. Nat Rev Immunol 11 : 403–415.

18. OhanianJ, OhanianV (2001) Sphingolipids in mammalian cell signalling. Cell Mol Life Sci 58 : 2053–2068.

19. van MeerG, VoelkerDR, FeigensonGW (2008) Membrane lipids: where they are and how they behave. Nat Rev Mol Cell Biol 9 : 112–124.

20. SchwabSR, PereiraJP, MatloubianM, XuY, HuangY, et al. (2005) Lymphocyte sequestration through S1P lyase inhibition and disruption of S1P gradients. Science 309 : 1735–1739.

21. VogelP, DonovielMS, ReadR, HansenGM, HazlewoodJ, et al. (2009) Incomplete inhibition of sphingosine 1-phosphate lyase modulates immune system function yet prevents early lethality and non-lymphoid lesions. PLoS One 4: e4112.

22. WalzerT, ChiossoneL, ChaixJ, CalverA, CarozzoC, et al. (2007) Natural killer cell trafficking in vivo requires a dedicated sphingosine 1-phosphate receptor. Nat Immunol 8 : 1337–1344.

23. WangF, Van BrocklynJR, HobsonJP, MovafaghS, Zukowska-GrojecZ, et al. (1999) Sphingosine 1-phosphate stimulates cell migration through a G(i)-coupled cell surface receptor. Potential involvement in angiogenesis. J Biol Chem 274 : 35343–35350.

24. PostmaFR, JalinkK, HengeveldT, MoolenaarWH (1996) Sphingosine-1-phosphate rapidly induces Rho-dependent neurite retraction: action through a specific cell surface receptor. EMBO J 15 : 2388–2392.

25. RoviezzoF, BrancaleoneV, De GruttolaL, VelleccoV, BucciM, et al. (2011) Sphingosine-1-phosphate modulates vascular permeability and cell recruitment in acute inflammation in vivo. J Pharmacol Exp Ther 337 : 830–837.

26. HammadSM (2011) Blood sphingolipids in homeostasis and pathobiology. Adv Exp Med Biol 721 : 57–66.

27. OskeritzianCA, PriceMM, HaitNC, KapitonovD, FalangaYT, et al. (2010) Essential roles of sphingosine-1-phosphate receptor 2 in human mast cell activation, anaphylaxis, and pulmonary edema. J Exp Med 207 : 465–474.

28. CamererE, RegardJB, CornelissenI, SrinivasanY, DuongDN, et al. (2009) Sphingosine-1-phosphate in the plasma compartment regulates basal and inflammation-induced vascular leak in mice. J Clin Invest 119 : 1871–1879.

29. JacksonAA, GrossMJ, DanielsEF, HamptonTH, HammondJH, et al. (2013) Anr and Its Activation by PlcH Activity in Pseudomonas aeruginosa Host Colonization and Virulence. J Bacteriol 195 : 3093–3104.

30. WargoMJ, SzwergoldBS, HoganDA (2008) Identification of two gene clusters and a transcriptional regulator required for Pseudomonas aeruginosa glycine betaine catabolism. J Bacteriol 190 : 2690–2699.

31. WargoMJ, HoTC, GrossMJ, WhittakerLA, HoganDA (2009) GbdR regulates Pseudomonas aeruginosa plcH and pchP transcription in response to choline catabolites. Infect Immun 77 : 1103–1111.

32. HampelKJ, LabauveAE, MeadowsJA, FitzsimmonsLF, NockAM, et al. (2013) Characterization of the GbdR regulon in Pseudomonas aeruginosa. J Bacteriol 196 : 7–15.

33. NelsonGJ (1967) Studies on the lipids of sheep red blood cells. I. Lipid composition in low and high potassium red cells. Lipids 2 : 64–71.

34. LengleE, GeyerRP (1972) Comparison of cellular lipids of serum-free strain L mouse fibroblasts. Biochim Biophys Acta 260 : 608–616.

35. YatomiY, IgarashiY, YangL, HisanoN, QiR, et al. (1997) Sphingosine 1-phosphate, a bioactive sphingolipid abundantly stored in platelets, is a normal constituent of human plasma and serum. J Biochem 121 : 969–973.

36. LawSLS, SquierCA, WertzPW (1994) Free Sphingosine in Oral Epithelium. Journal of Dental Research 73 : 108–108.

37. OkinoN, TaniM, ImayamaS, ItoM (1998) Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa. J Biol Chem 273 : 14368–14373.

38. LassmannT, SonnhammerEL (2005) Kalign–an accurate and fast multiple sequence alignment algorithm. BMC Bioinformatics 6 : 298.

39. WinsorGL, LamDK, FlemingL, LoR, WhitesideMD, et al. (2011) Pseudomonas Genome Database: improved comparative analysis and population genomics capability for Pseudomonas genomes. Nucleic Acids Res 39: D596–600.

40. LucchesiGI, LisaTA, DomenechCE (1989) Choline and betaine as inducer agents of Pseudomonas aeruginosa phospholipase C activity in high phosphate medium. FEMS Microbiol Lett 48 : 335–338.

41. SageAE, VasilAI, VasilML (1997) Molecular characterization of mutants affected in the osmoprotectant-dependent induction of phospholipase C in Pseudomonas aeruginosa PAO1. Mol Microbiol 23 : 43–56.

42. SageAE, VasilML (1997) Osmoprotectant-dependent expression of plcH, encoding the hemolytic phospholipase C, is subject to novel catabolite repression control in Pseudomonas aeruginosa PAO1. J Bacteriol 179 : 4874–4881.

43. ShortridgeVD, LazdunskiA, VasilML (1992) Osmoprotectants and phosphate regulate expression of phospholipase C in Pseudomonas aeruginosa. Mol Microbiol 6 : 863–871.

44. WargoMJ, HoganDA (2009) Identification of genes required for Pseudomonas aeruginosa carnitine catabolism. Microbiology 155 : 2411–2419.

45. FischerCL, DrakeD, DawsonDV, BlanchetteDR, BrogdenKA, et al. (2012) Antibacterial activity of sphingoid bases and fatty acids against gram-positive bacteria and gram-negative bacteria. Antimicrob Agents Chemother 56 : 1157–1161 Epub ahead of print.

46. TeichgraberV, UlrichM, EndlichN, RiethmullerJ, WilkerB, et al. (2008) Ceramide accumulation mediates inflammation, cell death and infection susceptibility in cystic fibrosis. Nat Med 14 : 382–391.

47. ParkK, EliasPM, ShinKO, LeeYM, HupeM, et al. (2013) A novel role of a lipid species, sphingosine-1-phosphate, in epithelial innate immunity. Mol Cell Biol 33 : 752–762.

48. GargSK, VolpeE, PalmieriG, MatteiM, GalatiD, et al. (2004) Sphingosine 1-phosphate induces antimicrobial activity both in vitro and in vivo. J Infect Dis 189 : 2129–2138.

49. MalikZA, ThompsonCR, HashimiS, PorterB, IyerSS, et al. (2003) Cutting edge: Mycobacterium tuberculosis blocks Ca2+ signaling and phagosome maturation in human macrophages via specific inhibition of sphingosine kinase. J Immunol 170 : 2811–2815.

50. GargSK, SantucciMB, PanittiM, PucilloL, BocchinoM, et al. (2006) Does sphingosine 1-phosphate play a protective role in the course of pulmonary tuberculosis? Clin Immunol 121 : 260–264.

51. CamachoLR, EnsergueixD, PerezE, GicquelB, GuilhotC (1999) Identification of a virulence gene cluster of Mycobacterium tuberculosis by signature-tagged transposon mutagenesis. Mol Microbiol 34 : 257–267.

52. RecchiC, SclaviB, RauzierJ, GicquelB, ReyratJM (2003) Mycobacterium tuberculosis Rv1395 is a class III transcriptional regulator of the AraC family involved in cytochrome P450 regulation. J Biol Chem 278 : 33763–33773.

53. GallegosMT, SchleifR, BairochA, HofmannK, RamosJL (1997) Arac/XylS family of transcriptional regulators. Microbiol Mol Biol Rev 61 : 393–410.

54. OkinoN, ItoM (2007) Ceramidase enhances phospholipase C-induced hemolysis by Pseudomonas aeruginosa. J Biol Chem 282 : 6021–6030.

55. ChatterjeeA, DuttaPK, ChowdhuryR (2007) Effect of fatty acids and cholesterol present in bile on expression of virulence factors and motility of Vibrio cholerae. Infect Immun 75 : 1946–1953.

56. LowdenMJ, SkorupskiK, PellegriniM, ChiorazzoMG, TaylorRK, et al. (2010) Structure of Vibrio cholerae ToxT reveals a mechanism for fatty acid regulation of virulence genes. Proc Natl Acad Sci U S A 107 : 2860–2865.

57. LubertoC, StonehouseMJ, CollinsEA, MarchesiniN, El-BawabS, et al. (2003) Purification, characterization, and identification of a sphingomyelin synthase from Pseudomonas aeruginosa. PlcH is a multifunctional enzyme. J Biol Chem 278 : 32733–32743.

58. RickettsCRSJ, TopleyE, LillyHA (1951) Human skin lipids with particular reference to the self-sterilising power of the skin . Clinical Science 10 : 89–111.

59. WertzPW, DowningDT (1990) Free sphingosine in human epidermis. J Invest Dermatol 94 : 159–161.

60. BibelDJ, AlyR, ShahS, ShinefieldHR (1993) Sphingosines: antimicrobial barriers of the skin. Acta Derm Venereol 73 : 407–411.

61. BibelDJ, AlyR, ShinefieldHR (1992) Antimicrobial activity of sphingosines. J Invest Dermatol 98 : 269–273.

62. FischerCL, DrakeDR, DawsonDV, BlanchetteDR, BrogdenKA, et al. (2012) Antibacterial activity of sphingoid bases and fatty acids against Gram-positive and Gram-negative bacteria. Antimicrob Agents Chemother 56 : 1157–1161.

63. FischerCL, WaltersKS, DrakeDR, BlanchetteDR, DawsonDV, et al. (2013) Sphingoid Bases Are Taken Up by Escherichia coli and Staphylococcus aureus and Induce Ultrastructural Damage. Skin Pharmacol Physiol 26 : 36–44.

64. KelleyLA, SternbergMJ (2009) Protein structure prediction on the Web: a case study using the Phyre server. Nat Protoc 4 : 363–371.

65. PotvinE, LehouxDE, Kukavica-IbruljI, RichardKL, SanschagrinF, et al. (2003) In vivo functional genomics of Pseudomonas aeruginosa for high-throughput screening of new virulence factors and antibacterial targets. Environ Microbiol 5 : 1294–1308.

66. NeidhardtFC, BlochPL, SmithDF (1974) Culture medium for enterobacteria. J Bacteriol 119 : 736–747.

67. LaBauveAE, WargoMJ (2012) Growth and laboratory maintenance of Pseudomonas aeruginosa. Curr Protoc Microbiol Chapter 6: Unit 6E 1.

68. WargoMJ, GrossMJ, RajamaniS, AllardJL, LundbladLK, et al. (2011) Hemolytic Phospholipase C Inhibition Protects Lung Function During Pseudomonas aeruginosa Infection. Am J Respir Crit Care Med 184 : 345–354.

69. WargoMJ (2013) Choline Catabolism to Glycine Betaine Contributes to Pseudomonas aeruginosa Survival during Murine Lung Infection. PLoS One 8: e56850.

70. ShanksRM, CaiazzaNC, HinsaSM, ToutainCM, O'TooleGA (2006) Saccharomyces cerevisiae-based molecular tool kit for manipulation of genes from gram-negative bacteria. Appl Environ Microbiol 72 : 5027–5036.

71. ChoiKH, SchweizerHP (2006) mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. Nat Protoc 1 : 153–161.

72. FitzsimmonsLF, HampelKJ, WargoMJ (2012) Cellular choline and glycine betaine pools impact osmoprotection and phospholipase C production in Pseudomonas aeruginosa. J Bacteriol 194 : 4718–4726.

73. Miller JH (1972) Experiments in molecular genetics. Cold Spring, NY: Cold Spring Harbor Laboratory.

74. WurtzelO, Yoder-HimesDR, HanK, DandekarAA, EdelheitS, et al. (2012) The single-nucleotide resolution transcriptome of Pseudomonas aeruginosa grown in body temperature. PLoS Pathog 8: e1002945.

75. JacobsMA, AlwoodA, ThaipisuttikulI, SpencerD, HaugenE, et al. (2003) Comprehensive transposon mutant library of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 100 : 14339–14344.

76. SimonR, PrieferU, PuhlerA (1983) A Broad Host Range Mobilization System for Invivo Genetic-Engineering -⁠ Transposon Mutagenesis in Gram-Negative Bacteria. Bio-Technology 1 : 784–791.