1887

Abstract

The evolution of antibiotic resistance in pathogenic bacteria is a growing global health problem which is gradually making the treatment of infectious diseases less efficient. Antimicrobial peptides are small charged molecules found in organisms from the complete phylogenetic spectrum. The peptides are attractive candidates for novel drug development due to their activity against bacteria that are resistant to conventional antibiotics, and reports of peptide resistance are rare in the clinical setting. Paradoxically, many clinically relevant bacteria have mechanisms that can recognize and respond to the presence of cationic antimicrobial peptides (CAMPs) in the environment by changing the properties of the microbial surface thereby increasing the tolerance of the microbes towards the peptides. In an essential component of this inducible tolerance mechanism is the lipopolysaccharide modification operon –PA3559 which encodes enzymes required for LPS alterations leading to increased antimicrobial peptide tolerance. The expression of the operon is induced by the presence of CAMPs in the environment but the molecular mechanisms underlying the cellular recognition of the peptides are poorly elucidated. In this work, we investigate the factors influencing expression by transposon mutagenesis and promoter green fluorescent protein reporters. We have identified a novel gene encoding a Mig-14-like protein that is required for recognition of the CAMPs colistin and Novispirin G10 by . Moreover, we show that this gene is also required for the formation of CAMP-tolerant subpopulations in hydrodynamic flow chamber biofilms.

Funding
This study was supported by the:
  • Danish Research Council
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2011-09-01
2024-03-28
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References

  1. Ausubel F. M., Brent M. R., Kingston R. E., Moore D. D., Siedman J. G., Smith J. A., Struhl K. ( 1996). Current Protocols in Molecular Biology New York: Wiley;
    [Google Scholar]
  2. Bader M. W., Sanowar S., Daley M. E., Schneider A. R., Cho U., Xu W., Klevit R. E., Le Moual H., Miller S. I. ( 2005). Recognition of antimicrobial peptides by a bacterial sensor kinase. Cell 122:461–472 [View Article][PubMed]
    [Google Scholar]
  3. Bailey J., Manoil C. ( 2002). Genome-wide internal tagging of bacterial exported proteins. Nat Biotechnol 20:839–842[PubMed] [CrossRef]
    [Google Scholar]
  4. Bollenbach T., Quan S., Chait R., Kishony R. ( 2009). Nonoptimal microbial response to antibiotics underlies suppressive drug interactions. Cell 139:707–718 [View Article][PubMed]
    [Google Scholar]
  5. Breazeale S. D., Ribeiro A. A., Raetz C. R. ( 2002). Oxidative decarboxylation of UDP-glucuronic acid in extracts of polymyxin-resistant Escherichia coli. Origin of lipid A species modified with 4-amino-4-deoxy-l-arabinose. J Biol Chem 277:2886–2896 [View Article][PubMed]
    [Google Scholar]
  6. Breazeale S. D., Ribeiro A. A., Raetz C. R. ( 2003). Origin of lipid A species modified with 4-amino-4-deoxy-l-arabinose in polymyxin-resistant mutants of Escherichia coli. An aminotransferase (ArnB) that generates UDP-4-deoxy-l-arabinose. J Biol Chem 278:24731–24739 [View Article][PubMed]
    [Google Scholar]
  7. Breukink E., Wiedemann I., van Kraaij C., Kuipers O. P., Sahl H., de Kruijff B. ( 1999). Use of the cell wall precursor lipid II by a pore-forming peptide antibiotic. Science 286:2361–2364 [View Article][PubMed]
    [Google Scholar]
  8. Brodsky I. E., Ernst R. K., Miller S. I., Falkow S. ( 2002). mig-14 is a Salmonella gene that plays a role in bacterial resistance to antimicrobial peptides. J Bacteriol 184:3203–3213 [View Article][PubMed]
    [Google Scholar]
  9. Brodsky I. E., Ghori N., Falkow S., Monack D. ( 2005). Mig-14 is an inner membrane-associated protein that promotes Salmonella typhimurium resistance to CRAMP, survival within activated macrophages and persistent infection. Mol Microbiol 55:954–972 [View Article][PubMed]
    [Google Scholar]
  10. Brogden K. A. ( 2005). Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria?. Nat Rev Microbiol 3:238–250 [View Article][PubMed]
    [Google Scholar]
  11. Brötz H., Bierbaum G., Leopold K., Reynolds P. E., Sahl H. G. ( 1998). The lantibiotic mersacidin inhibits peptidoglycan synthesis by targeting lipid II. Antimicrob Agents Chemother 42:154–160[PubMed]
    [Google Scholar]
  12. Ceri H., Olson M. E., Stremick C., Read R. R., Morck D., Buret A. ( 1999). The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms. J Clin Microbiol 37:1771–1776[PubMed]
    [Google Scholar]
  13. Choi K. H., Schweizer H. P. ( 2005). An improved method for rapid generation of unmarked Pseudomonas aeruginosa deletion mutants. BMC Microbiol 5:30 [View Article][PubMed]
    [Google Scholar]
  14. Christensen B. B., Sternberg C., Andersen J. B., Palmer R. J. Jr, Nielsen A. T., Givskov M., Molin S. ( 1999). Molecular tools for study of biofilm physiology. Methods Enzymol 310:20–42 [View Article][PubMed]
    [Google Scholar]
  15. Clark D. J., Maaloe O. ( 1967). DNA replication and division cycle in Escherichia coli . J Mol Biol 23:99–112 [View Article]
    [Google Scholar]
  16. Conly J., Johnston B. ( 2006). Colistin: the phoenix arises. Can J Infect Dis Med Microbiol 17:267–269[PubMed]
    [Google Scholar]
  17. Conrad R. S., Galanos C. ( 1989). Fatty acid alterations and polymyxin B binding by lipopolysaccharides from Pseudomonas aeruginosa adapted to polymyxin B resistance. Antimicrob Agents Chemother 33:1724–1728[PubMed] [CrossRef]
    [Google Scholar]
  18. Ernst R. K., Yi E. C., Guo L., Lim K. B., Burns J. L., Hackett M., Miller S. I. ( 1999). Specific lipopolysaccharide found in cystic fibrosis airway Pseudomonas aeruginosa . Science 286:1561–1565 [View Article][PubMed]
    [Google Scholar]
  19. Fernández L., Gooderham W. J., Bains M., McPhee J. B., Wiegand I., Hancock R. E. ( 2010). Adaptive resistance to the “last hope” antibiotics polymyxin B and colistin in Pseudomonas aeruginosa is mediated by the novel two-component regulatory system ParR–ParS. Antimicrob Agents Chemother 54:3372–3382 [View Article][PubMed]
    [Google Scholar]
  20. Folkesson A., Haagensen J. A., Zampaloni C., Sternberg C., Molin S. ( 2008). Biofilm induced tolerance towards antimicrobial peptides. PLoS ONE 3:e1891 [View Article][PubMed]
    [Google Scholar]
  21. Frederiksen B., Koch C., Høiby N. ( 1997). Antibiotic treatment of initial colonization with Pseudomonas aeruginosa postpones chronic infection and prevents deterioration of pulmonary function in cystic fibrosis. Pediatr Pulmonol 23:330–335 [View Article][PubMed]
    [Google Scholar]
  22. Gilleland H. E. Jr, Stinnett J. D., Eagon R. G. ( 1974). Ultrastructural and chemical alteration of the cell envelope of Pseudomonas aeruginosa, associated with resistance to ethylenediaminetetraacetate resulting from growth in a Mg2+-deficient medium. J Bacteriol 117:302–311[PubMed]
    [Google Scholar]
  23. Gunn J. S. ( 2001). Bacterial modification of LPS and resistance to antimicrobial peptides. J Endotoxin Res 7:57–62[PubMed] [CrossRef]
    [Google Scholar]
  24. Gunn J. S., Lim K. B., Krueger J., Kim K., Guo L., Hackett M., Miller S. I. ( 1998). PmrA–PmrB-regulated genes necessary for 4-aminoarabinose lipid A modification and polymyxin resistance. Mol Microbiol 27:1171–1182 [View Article][PubMed]
    [Google Scholar]
  25. Haagensen J. A., Klausen M., Ernst R. K., Miller S. I., Folkesson A., Tolker-Nielsen T., Molin S. ( 2007). Differentiation and distribution of colistin- and sodium dodecyl sulfate-tolerant cells in Pseudomonas aeruginosa biofilms. J Bacteriol 189:28–37 [View Article][PubMed]
    [Google Scholar]
  26. Hiemstra P. S., Fernie-King B. A., McMichael J., Lachmann P. J., Sallenave J. M. ( 2004). Antimicrobial peptides: mediators of innate immunity as templates for the development of novel anti-infective and immune therapeutics. Curr Pharm Des 10:2891–2905 [View Article][PubMed]
    [Google Scholar]
  27. Hoeprich P. D. ( 1970). The polymyxins. Med Clin North Am 54:1257–1265[PubMed]
    [Google Scholar]
  28. Høiby N., Krogh Johansen H., Moser C., Song Z., Ciofu O., Kharazmi A. ( 2001). Pseudomonas aeruginosa and the in vitro and in vivo biofilm mode of growth. Microbes Infect 3:23–35 [View Article][PubMed]
    [Google Scholar]
  29. Jacobs M. A., Alwood A., Thaipisuttikul I., Spencer D., Haugen E., Ernst S., Will O., Kaul R., Raymond C. et al. ( 2003). Comprehensive transposon mutant library of Pseudomonas aeruginosa . Proc Natl Acad Sci U S A 100:14339–14344 [View Article][PubMed]
    [Google Scholar]
  30. Khandelia H., Kaznessis Y. N. ( 2005). Molecular dynamics simulations of helical antimicrobial peptides in SDS micelles: what do point mutations achieve?. Peptides 26:2037–2049 [View Article][PubMed]
    [Google Scholar]
  31. King J. D., Kocíncová D., Westman E. L., Lam J. S. ( 2009). Review: lipopolysaccharide biosynthesis in Pseudomonas aeruginosa . Innate Immun 15:261–312 [View Article][PubMed]
    [Google Scholar]
  32. Koch B., Jensen L. E., Nybroe O. ( 2001). A panel of Tn7-based vectors for insertion of the GFP marker gene or for delivery of cloned DNA into Gram-negative bacteria at a neutral chromosomal site. J Microbiol Methods 45:187–195 [View Article][PubMed]
    [Google Scholar]
  33. Lam J., Matewish M., Poon K. K. H. ( 2004). Lipopolysaccharides of Pseudomonas aeruginosa . Pseudomonas vol. 3 Biosynthesis of Macromolecules and Molecular Metabolism341–361 Ramos J. L.
    [Google Scholar]
  34. Lambertsen L., Sternberg C., Molin S. ( 2004). Mini-Tn7 transposons for site-specific tagging of bacteria with fluorescent proteins. Environ Microbiol 6:726–732 [View Article][PubMed]
    [Google Scholar]
  35. Landman D., Georgescu C., Martin D. A., Quale J. ( 2008). Polymyxins revisited. Clin Microbiol Rev 21:449–465 [View Article][PubMed]
    [Google Scholar]
  36. Littlewood J. M., Miller M. G., Ghoneim A. T., Ramsden C. H. ( 1985). Nebulised colomycin for early pseudomonas colonisation in cystic fibrosis. Lancet 325:865 [View Article][PubMed]
    [Google Scholar]
  37. Manoil C. ( 2000). Tagging exported proteins using Escherichia coli alkaline phosphatase gene fusions. Methods Enzymol 326:35–47 [View Article][PubMed]
    [Google Scholar]
  38. Matewish M. ( 2004). The functional role of lipopolysaccharide in the cell envelope and surface proteins of Pseudomonas aeruginosa .
    [Google Scholar]
  39. McPhee J. B., Lewenza S., Hancock R. E. ( 2003). Cationic antimicrobial peptides activate a two-component regulatory system, PmrA–PmrB, that regulates resistance to polymyxin B and cationic antimicrobial peptides in Pseudomonas aeruginosa . Mol Microbiol 50:205–217 [View Article][PubMed]
    [Google Scholar]
  40. Moskowitz S. M., Ernst R. K., Miller S. I. ( 2004). PmrAB, a two-component regulatory system of Pseudomonas aeruginosa that modulates resistance to cationic antimicrobial peptides and addition of aminoarabinose to lipid A. J Bacteriol 186:575–579 [View Article][PubMed]
    [Google Scholar]
  41. Nickel J. C., Ruseska I., Wright J. B., Costerton J. W. ( 1985). Tobramycin resistance of Pseudomonas aeruginosa cells growing as a biofilm on urinary catheter material. Antimicrob Agents Chemother 27:619–624[PubMed] [CrossRef]
    [Google Scholar]
  42. Pamp S. J., Gjermansen M., Johansen H. K., Tolker-Nielsen T. ( 2008). Tolerance to the antimicrobial peptide colistin in Pseudomonas aeruginosa biofilms is linked to metabolically active cells, and depends on the pmr and mexABoprM genes. Mol Microbiol 68:223–240 [View Article][PubMed]
    [Google Scholar]
  43. Perron G. G., Zasloff M., Bell G. ( 2006). Experimental evolution of resistance to an antimicrobial peptide. Proc Biol Sci 273:251–256 [View Article][PubMed]
    [Google Scholar]
  44. Prince A. S. ( 2002). Biofilms, antimicrobial resistance, and airway infection. N Engl J Med 347:1110–1111 [View Article][PubMed]
    [Google Scholar]
  45. Sawai M. V., Waring A. J., Kearney W. R., McCray P. B. Jr, Forsyth W. R., Lehrer R. I., Tack B. F. ( 2002). Impact of single-residue mutations on the structure and function of ovispirin/novispirin antimicrobial peptides. Protein Eng 15:225–232 [View Article][PubMed]
    [Google Scholar]
  46. Schneider T., Kruse T., Wimmer R., Wiedemann I., Sass V., Pag U., Jansen A., Nielsen A. K., Mygind P. H. et al. ( 2010). Plectasin, a fungal defensin, targets the bacterial cell wall precursor Lipid II. Science 328:1168–1172 [View Article][PubMed]
    [Google Scholar]
  47. Southward C. M., Surette M. G. ( 2002). The dynamic microbe: green fluorescent protein brings bacteria to light. Mol Microbiol 45:1191–1196 [View Article][PubMed]
    [Google Scholar]
  48. Srinivas N., Jetter P., Ueberbacher B. J., Werneburg M., Zerbe K., Steinmann J., Van der Meijden B., Bernardini F., Lederer A. et al. ( 2010). Peptidomimetic antibiotics target outer-membrane biogenesis in Pseudomonas aeruginosa . Science 327:1010–1013 [View Article][PubMed]
    [Google Scholar]
  49. Steinstraesser L., Tack B. F., Waring A. J., Hong T., Boo L. M., Fan M. H., Remick D. I., Su G. L., Lehrer R. I., Wang S. C. ( 2002). Activity of novispirin G10 against Pseudomonas aeruginosa in vitro and in infected burns. Antimicrob Agents Chemother 46:1837–1844 [View Article][PubMed]
    [Google Scholar]
  50. Storm D. R., Rosenthal K. S., Swanson P. E. ( 1977). Polymyxin and related peptide antibiotics. Annu Rev Biochem 46:723–763 [View Article][PubMed]
    [Google Scholar]
  51. Travis S. M., Anderson N. N., Forsyth W. R., Espiritu C., Conway B. D., Greenberg E. P., McCray P. B. Jr, Lehrer R. I., Welsh M. J., Tack B. F. ( 2000). Bactericidal activity of mammalian cathelicidin-derived peptides. Infect Immun 68:2748–2755 [View Article][PubMed]
    [Google Scholar]
  52. Trent M. S., Ribeiro A. A., Doerrler W. T., Lin S., Cotter R. J., Raetz C. R. ( 2001). Accumulation of a polyisoprene-linked amino sugar in polymyxin-resistant Salmonella typhimurium and Escherichia coli: structural characterization and transfer to lipid A in the periplasm. J Biol Chem 276:43132–43144 [View Article][PubMed]
    [Google Scholar]
  53. Vaara M., Vaara T., Jensen M., Helander I., Nurminen M., Rietschel E. T., Mäkelä P. H. ( 1981). Characterization of the lipopolysaccharide from the polymyxin-resistant pmrA mutants of Salmonella typhimurium . FEBS Lett 129:145–149 [View Article][PubMed]
    [Google Scholar]
  54. Valdivia R. H., Cirillo D. M., Lee A. K., Bouley D. M., Falkow S. ( 2000). mig-14 is a horizontally acquired, host-induced gene required for Salmonella enterica lethal infection in the murine model of typhoid fever. Infect Immun 68:7126–7131 [View Article][PubMed]
    [Google Scholar]
  55. Winfield M. D., Groisman E. A. ( 2004). Phenotypic differences between Salmonella and Escherichia coli resulting from the disparate regulation of homologous genes. Proc Natl Acad Sci U S A 101:17162–17167 [View Article][PubMed]
    [Google Scholar]
  56. Winsor G. L., Van Rossum T., Lo R., Khaira B., Whiteside M. D., Hancock R. E., Brinkman F. S. ( 2009). Pseudomonas Genome Database: facilitating user-friendly, comprehensive comparisons of microbial genomes. Nucleic Acids Res 37:Database issueD483–D488 [View Article][PubMed]
    [Google Scholar]
  57. Zaslaver A., Bren A., Ronen M., Itzkovitz S., Kikoin I., Shavit S., Liebermeister W., Surette M. G., Alon U. ( 2006). A comprehensive library of fluorescent transcriptional reporters for Escherichia coli . Nat Methods 3:623–628 [View Article][PubMed]
    [Google Scholar]
  58. Zasloff M. ( 2002). Antimicrobial peptides of multicellular organisms. Nature 415:389–395 [View Article][PubMed]
    [Google Scholar]
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