1887

Abstract

Clinical isolates of that hyperproduce a dark-brown pigment are quite often found in the lungs of chronically infected patients, suggesting that they may have an adaptive advantage in chronic infections. We have screened a library of random transposon insertions in . Transposon insertions resulting in the hyperproduction of a dark-brown pigment were found to be located in the gene, which putatively encodes the enzyme homogentisate-1,2-dioxygenase. Complementation studies indicate that disruption is responsible for the hyperproduction of pyomelanin in both laboratory and clinical isolates. A relationship between disruption and adaptation to chronic infection was explored and our results show that the inactivation of produces a slight reduction of killing ability in an acute murine model of lung infection. On the other hand, it also confers decreased clearance and increased persistence in chronic lung infections. Whether pyomelanin production is the cause of the increased adaptation to chronicity or just a side effect of inactivation is a question to be studied in future; however, this adaptation is consistent with the higher resistance to oxidative stress conferred by the pyomelanin pigment. Our results clearly demonstrate that inactivation leads to a better adaptation to chronic infection, and strongly suggest that this mechanism may be exploited in naturally occurring strains.

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2009-04-01
2020-04-07
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References

  1. Arias-Barrau E., Olivera E. R., Luengo J. M., Fernandez C., Galan B., Garcia J. L., Diaz E., Minambres B.. 2004; The homogentisate pathway: a central catabolic pathway involved in the degradation of l-phenylalanine, l-tyrosine, and 3-hydroxyphenylacetate in Pseudomonas putida . J Bacteriol186:5062–5077
    [Google Scholar]
  2. Bodey G. P., Bolivar R., Fainstein V., Jadeja L.. 1983; Infections caused by Pseudomonas aeruginosa . Rev Infect Dis5:279–313
    [Google Scholar]
  3. Choi J. Y., Sifri C. D., Goumnerov B. C., Rahme L. G., Ausubel F. M., Calderwood S. B.. 2002; Identification of virulence genes in a pathogenic strain of Pseudomonas aeruginosa by representational difference analysis. J Bacteriol184:952–961
    [Google Scholar]
  4. Coon S. L., Kotob S., Jarvis B. B., Wang S., Fuqua W. C., Weiner R. M.. 1994; Homogentisic acid is the product of MelA, which mediates melanogenesis in the marine bacterium Shewanella colwelliana D. Appl Environ Microbiol60:3006–3010
    [Google Scholar]
  5. Doring G., Holder I. A., Botzenhardt K.. (editors) 1987; Basic Research and Clinical Aspects of Pseudomonas aeruginosa International Symposium; Tübingen, Germany: 1986; Basel: Karger;
  6. Driffield K., Miller K., Bostock J. M., O'Neill A. J., Chopra I.. 2008; Increased mutability of Pseudomonas aeruginosa in biofilms. J Antimicrob Chemother61:1053–1056
    [Google Scholar]
  7. Ernst R. K., D'Argenio D. A., Ichikawa J. K., Bangera M. G., Selgrade S., Burns J. L., Hiatt P., McCoy K., Brittnacher M., Kas A.. other authors 2003; Genome mosaicism is conserved but not unique in Pseudomonas aeruginosa isolates from the airways of young children with cystic fibrosis. Environ Microbiol5:1341–1349
    [Google Scholar]
  8. Fazal N.. 1997; The role of reactive oxygen species (ROS) in the effector mechanisms of human antimycobacterial immunity. Biochem Mol Biol Int43:399–408
    [Google Scholar]
  9. Fick R. B. Jr. 1993; Pathogenesis and disease. In Pseudomonas aeruginosa: the Opportunist Boca Raton, FL: CRC;
    [Google Scholar]
  10. Gilligan P. H.. 1991; Microbiology of airway disease in patients with cystic fibrosis. Clin Microbiol Rev4:35–51
    [Google Scholar]
  11. Govan J. R., Deretic V.. 1996; Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia . Microbiol Rev60:539–574
    [Google Scholar]
  12. Haussler S., Ziegler I., Lottel A., von Gotz F., Rohde M., Wehmhohner D., Saravanamuthu S., Tummler B., Steinmetz I.. 2003; Highly adherent small-colony variants of Pseudomonas aeruginosa in cystic fibrosis lung infection. J Med Microbiol52:295–301
    [Google Scholar]
  13. He J., Baldini R. L., Deziel E., Saucier M., Zhang Q., Liberati N. T., Lee D., Urbach J., Goodman H. M., Rahme L. G.. 2004; The broad host range pathogen Pseudomonas aeruginosa strain PA14 carries two pathogenicity islands harboring plant and animal virulence genes. Proc Natl Acad Sci U S A101:2530–2535
    [Google Scholar]
  14. Hoti S. L., Balaraman K.. 1993; Formation of melanin pigment by a mutant of Bacillus thuringiensis H-14. J Gen Microbiol139:2365–2369
    [Google Scholar]
  15. Hullo M. F., Moszer I., Danchin A., Martin-Verstraete I.. 2001; CotA of Bacillus subtilis is a copper-dependent laccase. J Bacteriol183:5426–5430
    [Google Scholar]
  16. Ivins B. E., Holmes R. K.. 1980; Isolation and characterization of melanin-producing ( mel ) mutants of Vibrio cholerae . Infect Immun27:721–729
    [Google Scholar]
  17. Ivins B. E., Holmes R. K.. 1981; Factors affecting phaeomelanin production by a melanin-producing ( mel ) mutant of Vibrio cholerae . Infect Immun34:895–899
    [Google Scholar]
  18. Jacobs M. A., Alwood A., Thaipisuttikul I., Spencer D., Haugen E., Ernst S., Will O., Kaul R., Raymond C.. other authors 2003; Comprehensive transposon mutant library of Pseudomonas aeruginosa . Proc Natl Acad Sci U S A100:14339–14344
    [Google Scholar]
  19. Johansen H. K., Espersen F., Cryz S. J. Jr, Hougen H. P., Fomsgaard A., Rygaard J., Hoiby N.. 1994; Immunization with Pseudomonas aeruginosa vaccines and adjuvant can modulate the type of inflammatory response subsequent to infection. Infect Immun62:3146–3155
    [Google Scholar]
  20. Konstan M. W., Hilliard K. A., Norvell T. M., Berger M.. 1994; Bronchoalveolar lavage findings in cystic fibrosis patients with stable, clinically mild lung disease suggest ongoing infection and inflammation. Am J Respir Crit Care Med150:448–454
    [Google Scholar]
  21. Kotob S. I., Coon S. L., Quintero E. J., Weiner R. M.. 1995; Homogentisic acid is the primary precursor of melanin synthesis in Vibrio cholerae , a Hyphomonas strain, and Shewanella colwelliana . Appl Environ Microbiol61:1620–1622
    [Google Scholar]
  22. Kovach M. E., Elzer P. H., Hill D. S., Robertson G. T., Farris M. A., Roop R. M. II, Peterson K. M.. 1995; Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene166:175–176
    [Google Scholar]
  23. Liberati N. T., Urbach J. M., Miyata S., Lee D. G., Drenkard E., Wu G., Villanueva J., Wei T., Ausubel F. M.. 2006; An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants. Proc Natl Acad Sci U S A103:2833–2838
    [Google Scholar]
  24. Macia M. D., Borrell N., Segura M., Gomez C., Perez J. L., Oliver A.. 2006; Efficacy and potential for resistance selection of antipseudomonal treatments in a mouse model of lung infection by hypermutable Pseudomonas aeruginosa . Antimicrob Agents Chemother50:975–983
    [Google Scholar]
  25. Mena A., Macia M. D., Borrell N., Moya B., de Francisco T., Perez J. L., Oliver A.. 2007; Inactivation of the mismatch repair system in Pseudomonas aeruginosa attenuates virulence but favors persistence of oropharyngeal colonization in cystic fibrosis mice. J Bacteriol189:3665–3668
    [Google Scholar]
  26. Morrison A. J. Jr, Wenzel R. P.. 1984; Epidemiology of infections due to Pseudomonas aeruginosa . Rev Infect Dis6:Suppl. 3S627–S642
    [Google Scholar]
  27. Mulcahy H., O'Callaghan J., O'Grady E. P., Maciá M. D., Borrell N., Gómez C., Casey P. G., Hill C., Adams C.. other authors 2008; Pseudomonas aeruginosa RsmA plays an important role during murine infection by influencing colonization, virulence, persistence, and pulmonary inflammation. Infect Immun76:632–638
    [Google Scholar]
  28. Nosanchuk J. D., Casadevall A.. 2003; The contribution of melanin to microbial pathogenesis. Cell Microbiol5:203–223
    [Google Scholar]
  29. Nosanchuk J. D., Casadevall A.. 2006; Impact of melanin on microbial virulence and clinical resistance to antimicrobial compounds. Antimicrob Agents Chemother50:3519–3528
    [Google Scholar]
  30. Oliver A., Canton R., Campo P., Baquero F., Blazquez J.. 2000; High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science288:1251–1254
    [Google Scholar]
  31. Pritt B., O'Brien L., Winn W.. 2007; Mucoid Pseudomonas in cystic fibrosis. Am J Clin Pathol128:32–34
    [Google Scholar]
  32. Ruzafa C., Sanchez-Amat A., Solano F.. 1995; Characterization of the melanogenic system in Vibrio cholerae , ATCC 14035. Pigment Cell Res8:147–152
    [Google Scholar]
  33. Salgado A., Blazquez J.. 2006; Determination of the structure of a hybrid between 2-(1,4-benzoquinone)acetic acid and a linear peptide by electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom20:512–516
    [Google Scholar]
  34. Sambrook J., Russell D. W.. 2001; Molecular Cloning: a Laboratory Manual , 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  35. Smith E. E., Buckley D. G., Wu Z., Saenphimmachak C., Hoffman L. R., D'Argenio D. A., Miller S. I., Ramsey B. W., Speert D. P.. other authors 2006; Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients. Proc Natl Acad Sci U S A103:8487–8492
    [Google Scholar]
  36. Stover C. K., Pham X. Q., Erwin A. L., Mizoguchi S. D., Warrener P., Hickey M. J., Brinkman F. S., Hufnagle W. O., Kowalik D. J.. other authors 2000; Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature406:959–964
    [Google Scholar]
  37. Yabuuchi E., Ohyama A.. 1972; Characterization of pyomelanin producing strains of Pseudomonas aeruginosa . Int J Syst Bacteriol22:53–64
    [Google Scholar]
  38. Zhang C., Huang M., Holloway B. W.. 1993; Mapping of the ben , ant and cat genes of Pseudomonas aeruginosa and evolutionary relationship of the ben region of P. aeruginosa and P. putida . FEMS Microbiol Lett108:303–309
    [Google Scholar]
  39. Zughaier S. M., Ryley H. C., Jackson S. K.. 1999; A melanin pigment purified from an epidemic strain of Burkholderia cepacia attenuates monocyte respiratory burst activity by scavenging superoxide anion. Infect Immun67:908–913
    [Google Scholar]
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