1887

Abstract

is an opportunistic pathogen that in the environment colonizes biofilms and replicates within amoebae. The bacteria employ the intracellular multiplication/defective organelle trafficking (Icm/Dot) type IV secretion system to grow intracellularly in a specific vacuole. Using as a host cell, we have previously identified ( cytotoxic suppressor), a paralogue of the lipid A disaccharide synthase , as a cytotoxic factor of . A bioinformatic analysis of the genome revealed that is unique in harbouring two paralogues of and two and three paralogues of the lipid A biosynthesis acyltransferases and , respectively. () forms a transcriptional unit with , encoding a putative UDP-GlcNAc oxidase in the biosynthetic pathway leading to 3-aminoglucosamine analogues of lipid A. clusters with , and paralogues, encoding secondary acyltransferases. / and were found to partially complement the growth defect of an conditional mutant strain, indicating that both corresponding enzymes possess lipid A disaccharide synthase activity. The two paralogues are not functionally equivalent, since expression of / but not in an mutant is cytotoxic for , and LPS purified from the two strains triggers CD14-dependent tumour necrosis factor (TNF) production by macrophages with a different potency. The and paralogues are expressed under various growth conditions, including broth, biofilms and in . While the flagellar gene is mainly expressed in late stationary phase, the and paralogues are preferentially expressed in the exponential and early stationary phases. Upon exposure to hypotonic stress and nutrient deprivation, , and to a lesser extent /, is upregulated. The differential regulation of or paralogues in response to changing environmental conditions might allow to adapt its lipid A structure.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2007/009829-0
2007-11-01
2019-11-15
Loading full text...

Full text loading...

/deliver/fulltext/micro/153/11/3817.html?itemId=/content/journal/micro/10.1099/mic.0.2007/009829-0&mimeType=html&fmt=ahah

References

  1. Albers, U., Reus, K., Shuman, H. A. & Hilbi, H. ( 2005; ). The amoebae plate test implicates a paralogue of lpxB in the interaction of Legionella pneumophila with Acanthamoeba castellanii. Microbiology 151, 167–182.[CrossRef]
    [Google Scholar]
  2. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997; ). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.[CrossRef]
    [Google Scholar]
  3. Barker, J., Lambert, P. A. & Brown, M. R. ( 1993; ). Influence of intra-amoebic and other growth conditions on the surface properties of Legionella pneumophila. Infect Immun 61, 3503–3510.
    [Google Scholar]
  4. Bishop, R. E., Gibbons, H. S., Guina, T., Trent, M. S., Miller, S. I. & Raetz, C. R. ( 2000; ). Transfer of palmitate from phospholipids to lipid A in outer membranes of Gram-negative bacteria. EMBO J 19, 5071–5080.[CrossRef]
    [Google Scholar]
  5. Brüggemann, H., Cazalet, C. & Buchrieser, C. ( 2006; ). Adaptation of Legionella pneumophila to the host environment: role of protein secretion, effectors and eukaryotic-like proteins. Curr Opin Microbiol 9, 86–94.[CrossRef]
    [Google Scholar]
  6. Byrne, B. & Swanson, M. S. ( 1998; ). Expression of Legionella pneumophila virulence traits in response to growth conditions. Infect Immun 66, 3029–3034.
    [Google Scholar]
  7. Cazalet, C., Rusniok, C., Brüggemann, H., Zidane, N., Magnier, A., Ma, L., Tichit, M., Jarraud, S., Bouchier, C. & other authors ( 2004; ). Evidence in the Legionella pneumophila genome for exploitation of host cell functions and high genome plasticity. Nat Genet 36, 1165–1173.[CrossRef]
    [Google Scholar]
  8. Chien, M., Morozova, I., Shi, S., Sheng, H. & Chen J Gomez, S. M., Asamani, G., Hill, K., Nuara, J. & other authors ( 2004; ). The genomic sequence of the accidental pathogen Legionella pneumophila. Science 305, 1966–1968.[CrossRef]
    [Google Scholar]
  9. Crowell, D. N., Anderson, M. S. & Raetz, C. R. ( 1986; ). Molecular cloning of the genes for lipid A disaccharide synthase and UDP-N-acetylglucosamine acyltransferase in Escherichia coli. J Bacteriol 168, 152–159.
    [Google Scholar]
  10. Crowell, D. N., Reznikoff, W. S. & Raetz, C. R. ( 1987; ). Nucleotide sequence of the Escherichia coli gene for lipid A disaccharide synthase. J Bacteriol 169, 5727–5734.
    [Google Scholar]
  11. Feeley, J. C., Gibson, R. J., Gorman, G. W., Langford, N. C., Rasheed, J. K., Mackel, D. C. & Baine, W. B. ( 1979; ). Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila. J Clin Microbiol 10, 437–441.
    [Google Scholar]
  12. Fernandez-Moreira, E., Helbig, J. H. & Swanson, M. S. ( 2006; ). Membrane vesicles shed by Legionella pneumophila inhibit fusion of phagosomes with lysosomes. Infect Immun 74, 3285–3295.[CrossRef]
    [Google Scholar]
  13. Fields, B. S. ( 1996; ). The molecular ecology of legionellae. Trends Microbiol 4, 286–290.[CrossRef]
    [Google Scholar]
  14. Gangloff, S. C., Hijiya, N., Haziot, A. & Goyert, S. M. ( 1999; ). Lipopolysaccharide structure influences the macrophage response via CD14-independent and CD14-dependent pathways. Clin Infect Dis 28, 491–496.[CrossRef]
    [Google Scholar]
  15. Gangloff, S. C., Zähringer, U., Blondin, C., Guenounou, M., Silver, J. & Goyert, S. M. ( 2005; ). Influence of CD14 on ligand interactions between lipopolysaccharide and its receptor complex. J Immunol 175, 3940–3945.[CrossRef]
    [Google Scholar]
  16. Girard, R., Pedron, T., Uematsu, S., Balloy, V., Chignard, M., Akira, S. & Chaby, R. ( 2003; ). Lipopolysaccharides from Legionella and Rhizobium stimulate mouse bone marrow granulocytes via Toll-like receptor 2. J Cell Sci 116, 293–302.[CrossRef]
    [Google Scholar]
  17. Guo, L., Lim, K. B., Poduje, C. M., Daniel, M., Gunn, J. S., Hackett, M. & Miller, S. I. ( 1998; ). Lipid A acylation and bacterial resistance against vertebrate antimicrobial peptides. Cell 95, 189–198.[CrossRef]
    [Google Scholar]
  18. Haziot, A., Ferrero, E., Kontgen, F., Hijiya, N., Yamamoto, S., Silver, J., Stewart, C. L. & Goyert, S. M. ( 1996; ). Resistance to endotoxin shock and reduced dissemination of Gram-negative bacteria in CD14-deficient mice. Immunity 4, 407–414.[CrossRef]
    [Google Scholar]
  19. Heuner, K., Brand, B. C. & Hacker, J. ( 1999; ). The expression of the flagellum of Legionella pneumophila is modulated by different environmental factors. FEMS Microbiol Lett 175, 69–77.[CrossRef]
    [Google Scholar]
  20. Hilbi, H. ( 2006; ). Modulation of phosphoinositide metabolism by pathogenic bacteria. Cell Microbiol 8, 1697–1706.[CrossRef]
    [Google Scholar]
  21. Hilbi, H., Segal, G. & Shuman, H. A. ( 2001; ). Icm/Dot-dependent upregulation of phagocytosis by Legionella pneumophila. Mol Microbiol 42, 603–617.
    [Google Scholar]
  22. Hilbi, H., Weber, S. S., Ragaz, C., Nyfeler, Y. & Urwyler, S. ( 2007; ). Environmental predators as models for bacterial pathogenesis. Environ Microbiol 9, 563–575.[CrossRef]
    [Google Scholar]
  23. Jürgens, D. & Fehrenbach, F. J. ( 1997; ). Identification of Legionella species by lipopolysaccharide antigen pattern. J Clin Microbiol 35, 3054–3057.
    [Google Scholar]
  24. Knirel, Y. A., Rietschel, E. T., Marre, R. & Zähringer, U. ( 1994; ). The structure of the O-specific chain of Legionella pneumophila serogroup 1 lipopolysaccharide. Eur J Biochem 221, 239–245.[CrossRef]
    [Google Scholar]
  25. Kooistra, O., Knirel, Y. A., Lüneberg, E., Frosch, M. & Zähringer, U. ( 2002a; ). Phase variation in Legionella pneumophila serogroup 1, subgroup OLDA, strain RC1 influences lipid A structure. In: Legionella – Proceedings of the 5th International Conference, pp. 68–73. Edited by R. Marre, Y. Abu Kwaik, C. Bartlett, N. Cianciotto, B. S. Fields, M. Frosch, J. Hacker & B. C. Lueck. Washington, DC: ASM Press.
  26. Kooistra, O., Lüneberg, E., Knirel, Y. A., Frosch, M. & Zähringer, U. ( 2002b; ). N-Methylation in polylegionaminic acid is associated with the phase-variable epitope of Legionella pneumophila serogroup 1 lipopolysaccharide. Identification of 5-(N,N-dimethylacetimidoyl) amino and 5-acetimidoyl(N-methyl)amino-7-acetamido-3,5,7,9-tetradeoxynon-2-ulosonic acid in the O-chain polysaccharide. Eur J Biochem 269, 560–572.[CrossRef]
    [Google Scholar]
  27. Lüneberg, E., Zähringer, U., Knirel, Y. A., Steinmann, D., Hartmann, M., Steinmetz, I., Rohde, M., Kohl, J. & Frosch, M. ( 1998; ). Phase-variable expression of lipopolysaccharide contributes to the virulence of Legionella pneumophila. J Exp Med 188, 49–60.[CrossRef]
    [Google Scholar]
  28. Lüneberg, E., Mayer, B., Daryab, N., Kooistra, O., Zähringer, U., Rohde, M., Swanson, J. & Frosch, M. ( 2001; ). Chromosomal insertion and excision of a 30 kb unstable genetic element is responsible for phase variation of lipopolysaccharide and other virulence determinants in Legionella pneumophila. Mol Microbiol 39, 1259–1271.[CrossRef]
    [Google Scholar]
  29. Mampel, J., Spirig, T., Weber, S. S., Haagensen, J. A., Molin, S. & Hilbi, H. ( 2006; ). Planktonic replication is essential for biofilm formation by Legionella pneumophila in a complex medium under static and dynamic flow conditions. Appl Environ Microbiol 72, 2885–2895.[CrossRef]
    [Google Scholar]
  30. Manthey, C. L., Perera, P. Y., Henricson, B. E., Hamilton, T. A., Qureshi, N. & Vogel, S. N. ( 1994; ). Endotoxin-induced early gene expression in C3H/HeJ (Lpsd) macrophages. J Immunol 153, 2653–2663.
    [Google Scholar]
  31. Miller, S. I., Ernst, R. K. & Bader, M. W. ( 2005; ). LPS, TLR4 and infectious disease diversity. Nat Rev Microbiol 3, 36–46.[CrossRef]
    [Google Scholar]
  32. Moffat, J. F. & Tompkins, L. S. ( 1992; ). A quantitative model of intracellular growth of Legionella pneumophila in Acanthamoeba castellanii. Infect Immun 60, 296–301.
    [Google Scholar]
  33. Moll, H., Sonesson, A., Jantzen, E., Marre, R. & Zähringer, U. ( 1992; ). Identification of 27-oxo-octacosanoic acid and heptacosane-1,27-dioic acid in Legionella pneumophila. FEMS Microbiol Lett 76, 1–6.
    [Google Scholar]
  34. Molofsky, A. B. & Swanson, M. S. ( 2004; ). Differentiate to thrive: lessons from the Legionella pneumophila life cycle. Mol Microbiol 53, 29–40.[CrossRef]
    [Google Scholar]
  35. Murga, R., Forster, T. S., Brown, E., Pruckler, J. M., Fields, B. S. & Donlan, R. M. ( 2001; ). Role of biofilms in the survival of Legionella pneumophila in a model potable-water system. Microbiology 147, 3121–3126.
    [Google Scholar]
  36. Nagai, H. & Roy, C. R. ( 2003; ). Show me the substrates: modulation of host cell function by type IV secretion systems. Cell Microbiol 5, 373–383.[CrossRef]
    [Google Scholar]
  37. Nash, T. W., Libby, D. M. & Horwitz, M. A. ( 1984; ). Interaction between the Legionnaires' disease bacterium (Legionella pneumophila) and human alveolar macrophages. Influence of antibody, lymphokines, and hydrocortisone. J Clin Invest 74, 771–782.[CrossRef]
    [Google Scholar]
  38. Neumeister, B., Faigle, M., Sommer, M., Zähringer, U., Stelter, F., Menzel, R., Schutt, C. & Northoff, H. ( 1998; ). Low endotoxic potential of Legionella pneumophila lipopolysaccharide due to failure of interaction with the monocyte lipopolysaccharide receptor CD14. Infect Immun 66, 4151–4157.
    [Google Scholar]
  39. Nishijima, M., Bulawa, C. E. & Raetz, C. R. ( 1981; ). Two interacting mutations causing temperature-sensitive phosphatidylglycerol synthesis in Escherichia coli membranes. J Bacteriol 145, 113–121.
    [Google Scholar]
  40. Raetz, C. R. & Whitfield, C. ( 2002; ). Lipopolysaccharide endotoxins. Annu Rev Biochem 71, 635–700.[CrossRef]
    [Google Scholar]
  41. Robey, M., O'Connell, W. & Cianciotto, N. P. ( 2001; ). Identification of Legionella pneumophila rcp, a pagP-like gene that confers resistance to cationic antimicrobial peptides and promotes intracellular infection. Infect Immun 69, 4276–4286.[CrossRef]
    [Google Scholar]
  42. Segal, G. & Shuman, H. A. ( 1999; ). Legionella pneumophila utilizes the same genes to multiply within Acanthamoeba castellanii and human macrophages. Infect Immun 67, 2117–2124.
    [Google Scholar]
  43. Segal, G., Purcell, M. & Shuman, H. A. ( 1998; ). Host cell killing and bacterial conjugation require overlapping sets of genes within a 22-kb region of the Legionella pneumophila genome. Proc Natl Acad Sci U S A 95, 1669–1674.[CrossRef]
    [Google Scholar]
  44. Spörri, R., Joller, N., Albers, U., Hilbi, H. & Oxenius, A. ( 2006; ). MyD88-dependent IFN-γ production by NK cells is key for control of Legionella pneumophila infection. J Immunol 176, 6162–6171.[CrossRef]
    [Google Scholar]
  45. Steinert, M., Hentschel, U. & Hacker, J. ( 2002; ). Legionella pneumophila: an aquatic microbe goes astray. FEMS Microbiol Rev 26, 149–162.[CrossRef]
    [Google Scholar]
  46. Sweet, C. R., Ribeiro, A. A. & Raetz, C. R. ( 2004; ). Oxidation and transamination of the 3′′-position of UDP-N-acetylglucosamine by enzymes from Acidithiobacillus ferrooxidans. Role in the formation of lipid A molecules with four amide-linked acyl chains. J Biol Chem 279, 25400–25410.[CrossRef]
    [Google Scholar]
  47. Tiaden, A., Spirig, T., Weber, S. S., Brüggemann, H., Bosshard, R., Buchrieser, C. & Hilbi, H. ( 2007; ). The Legionella pneumophila response regulator LqsR promotes host cell interactions as an element of the virulence regulatory network controlled by RpoS and LetA. Cell Microbiol
    [Google Scholar]
  48. Vogel, J. P., Andrews, H. L., Wong, S. K. & Isberg, R. R. ( 1998; ). Conjugative transfer by the virulence system of Legionella pneumophila. Science 279, 873–876.[CrossRef]
    [Google Scholar]
  49. Wiater, L. A., Sadosky, A. B. & Shuman, H. A. ( 1994; ). Mutagenesis of Legionella pneumophila using Tn903dlllacZ: identification of a growth-phase-regulated pigmentation gene. Mol Microbiol 11, 641–653.[CrossRef]
    [Google Scholar]
  50. Yamamoto, Y., Klein, T. W., Newton, C. A., Widen, R. & Friedman, H. ( 1988; ). Growth of Legionella pneumophila in thioglycolate-elicited peritoneal macrophages from A/J mice. Infect Immun 56, 370–375.
    [Google Scholar]
  51. Zähringer, U., Knirel, Y. A., Lindner, B., Helbig, J. H., Sonesson, A., Marre, R. & Rietschel, E. T. ( 1995; ). The lipopolysaccharide of Legionella pneumophila serogroup 1 (strain Philadelphia 1): chemical structure and biological significance. Prog Clin Biol Res 392, 113–139.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2007/009829-0
Loading
/content/journal/micro/10.1099/mic.0.2007/009829-0
Loading

Data & Media loading...

Supplements

Supplementary Fig. S1 legend [PDF file](13 KB)

PDF

Supplementary Tables [PDF file](81 KB)

PDF

Supplementary Fig. S1 [PDF file](118 KB)

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error