1887

Abstract

In common with other members of the complex (BCC), is capable of producing exopolysaccharide (EPS) when grown on certain mannitol-rich media. The significance of the resulting mucoid phenotype and the genome-wide response to mannitol has never been characterized despite its clinical relevance following the approval of a dried-powder preparation of mannitol as an inhaled osmolyte therapy for cystic fibrosis (CF) patients. In the present study we defined the transcriptional response of ATCC 17616, a model genome-sequenced strain of environmental origin, to growth on mannitol-rich yeast extract media (MYEM). EPS-dependent and -independent impact of MYEM on virulence-associated traits was assessed in both strain ATCC 17616 and the CF isolate C1576. Our studies revealed a significant transcriptional response to MYEM encompassing approximately 23 % of predicted genes within the genome. Strikingly, this transcriptional response identified that EPS induction occurs in ATCC 17616 without the upregulation of the and EPS gene clusters, despite their pivotal role in EPS biosynthesis. Of approximately 20 differentially expressed putative virulence factors, 16 exhibited upregulation including flagella, ornibactin, oxidative stress proteins and phospholipases. MYEM-grown also exhibited enhanced motility, biofilm formation and epithelial cell invasion. In contrast to these potential virulence enhancements, MYEM-grown C1576 showed attenuated virulence in the infection model. All of the observed phenotypic responses occurred independently of EPS production, highlighting the profound impact that mannitol-based growth has on the physiology and virulence of .

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2014-01-01
2020-01-22
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References

  1. Allenza P., Lee Y. N., Lessie T. G..( 1982;). Enzymes related to fructose utilization in Pseudomonas cepacia.. J Bacteriol150:1348–1356[PubMed]
    [Google Scholar]
  2. Allenza P., Morrell M. J., Detroy R. W..( 1990;). Conversion of mannose to fructose by immobilized mannose isomerase from Pseudomonas cepacia.. Appl Biochem Biotechnol24-25:171–182 [CrossRef][PubMed]
    [Google Scholar]
  3. Allison K. R., Brynildsen M. P., Collins J. J..( 2011;). Metabolite-enabled eradication of bacterial persisters by aminoglycosides. Nature473:216–220 [CrossRef][PubMed]
    [Google Scholar]
  4. Baran D., de Vuyst P., Ooms H. A..( 1990;). Concentration of tobramycin given by aerosol in the fluid obtained by bronchoalveolar lavage. Respir Med84:203–204 [CrossRef][PubMed]
    [Google Scholar]
  5. Bartholdson S. J., Brown A. R., Mewburn B. R., Clarke D. J., Fry S. C., Campopiano D. J., Govan J. R..( 2008;). Plant host and sugar alcohol induced exopolysaccharide biosynthesis in the Burkholderia cepacia complex. Microbiology154:2513–2521 [CrossRef][PubMed]
    [Google Scholar]
  6. Bilton D., Robinson P., Cooper P., Gallagher C. G., Kolbe J., Fox H., Jaques A., Charlton B..CF301 Study Investigators( 2011;). Inhaled dry powder mannitol in cystic fibrosis: an efficacy and safety study. Eur Respir J38:1071–1080 [CrossRef][PubMed]
    [Google Scholar]
  7. Brünker P., Altenbuchner J., Mattes R..( 1998;). Structure and function of the genes involved in mannitol, arabitol and glucitol utilization from Pseudomonas fluorescens DSM50106. Gene206:117–126 [CrossRef][PubMed]
    [Google Scholar]
  8. Bucior I., Abbott J., Song Y., Matthay M. A., Engel J. N..( 2013;). Sugar administration is an effective adjunctive therapy in the treatment of Pseudomonas aeruginosa pneumonia. Am J Physiol Lung Cell Mol Physiol305:L352–L363 [CrossRef][PubMed]
    [Google Scholar]
  9. Burns J. L., Jonas M., Chi E. Y., Clark D. K., Berger A., Griffith A..( 1996;). Invasion of respiratory epithelial cells by Burkholderia (Pseudomonas) cepacia.. Infect Immun64:4054–4059[PubMed]
    [Google Scholar]
  10. Bylund J., Burgess L. A., Cescutti P., Ernst R. K., Speert D. P..( 2006;). Exopolysaccharides from Burkholderia cenocepacia inhibit neutrophil chemotaxis and scavenge reactive oxygen species. J Biol Chem281:2526–2532 [CrossRef][PubMed]
    [Google Scholar]
  11. Conway B. A., Chu K. K., Bylund J., Altman E., Speert D. P..( 2004;). Production of exopolysaccharide by Burkholderia cenocepacia results in altered cell-surface interactions and altered bacterial clearance in mice. J Infect Dis190:957–966 [CrossRef][PubMed]
    [Google Scholar]
  12. Courtney J. M., Dunbar K. E., McDowell A., Moore J. E., Warke T. J., Stevenson M., Elborn J. S..( 2004;). Clinical outcome of Burkholderia cepacia complex infection in cystic fibrosis adults. J Cyst Fibros3:93–98 [CrossRef][PubMed]
    [Google Scholar]
  13. Cunha M. V., Sousa S. A., Leitão J. H., Moreira L. M., Videira P. A., Sá-Correia I..( 2004;). Studies on the involvement of the exopolysaccharide produced by cystic fibrosis-associated isolates of the Burkholderia cepacia complex in biofilm formation and in persistence of respiratory infections. J Clin Microbiol42:3052–3058 [CrossRef][PubMed]
    [Google Scholar]
  14. Daviskas E., Anderson S. D., Jaques A., Charlton B..( 2010;). Inhaled mannitol improves the hydration and surface properties of sputum in patients with cystic fibrosis. Chest137:861–868 [CrossRef][PubMed]
    [Google Scholar]
  15. Denman C. C., Brown A. R..( 2013;). Mannitol promotes adherence of an outbreak strain of Burkholderia multivorans via an exopolysaccharide-independent mechanism that is associated with upregulation of newly identified fimbrial and afimbrial adhesins. Microbiology159:771–781 [CrossRef][PubMed]
    [Google Scholar]
  16. Dubois M., Gilles K., Hamilton J. K., Rebers P. A., Smith F..( 1951;). A colorimetric method for the determination of sugars. Nature168:167 [CrossRef][PubMed]
    [Google Scholar]
  17. Etherington C., Peckham D. G., Conway S. P., Denton M..( 2003;). Burkholderia cepacia complex infection in adult patients with cystic fibrosis—is early eradication possible. J Cyst Fibros2:220–221 [CrossRef][PubMed]
    [Google Scholar]
  18. Ferreira A. S., Leitão J. H., Sousa S. A., Cosme A. M., Sá-Correia I., Moreira L. M..( 2007;). Functional analysis of Burkholderia cepacia genes bceD and bceF, encoding a phosphotyrosine phosphatase and a tyrosine autokinase, respectively: role in exopolysaccharide biosynthesis and biofilm formation. Appl Environ Microbiol73:524–534 [CrossRef][PubMed]
    [Google Scholar]
  19. Ferreira A. S., Leitão J. H., Silva I. N., Pinheiro P. F., Sousa S. A., Ramos C. G., Moreira L. M..( 2010;). Distribution of cepacian biosynthesis genes among environmental and clinical Burkholderia strains and role of cepacian exopolysaccharide in resistance to stress conditions. Appl Environ Microbiol76:441–450 [CrossRef][PubMed]
    [Google Scholar]
  20. Ferreira A. S., Silva I. N., Oliveira V. H., Becker J. D., Givskov M., Ryan R. P., Fernandes F., Moreira L. M..( 2013;). Comparative transcriptomic analysis of the Burkholderia cepacia tyrosine kinase bceF mutant reveals a role in tolerance to stress, biofilm formation, and virulence. Appl Environ Microbiol79:3009–3020 [CrossRef][PubMed]
    [Google Scholar]
  21. Flannagan R. S., Aubert D., Kooi C., Sokol P. A., Valvano M. A..( 2007;). Burkholderia cenocepacia requires a periplasmic HtrA protease for growth under thermal and osmotic stress and for survival in vivo. Infect Immun75:1679–1689 [CrossRef][PubMed]
    [Google Scholar]
  22. Gotschlich A., Huber B., Geisenberger O., Tögl A., Steidle A., Riedel K., Hill P., Tümmler B., Vandamme P..& other authors ( 2001;). Synthesis of multiple N-acylhomoserine lactones is wide-spread among the members of the Burkholderia cepacia complex. Syst Appl Microbiol24:1–14 [CrossRef][PubMed]
    [Google Scholar]
  23. Govan J. R., Brown A. R., Jones A. M..( 2007;). Evolving epidemiology of Pseudomonas aeruginosa and the Burkholderia cepacia complex in cystic fibrosis lung infection. Future Microbiol2:153–164 [CrossRef][PubMed]
    [Google Scholar]
  24. Graindorge A., Menard A., Neto M., Bouvet C., Miollan R., Gaillard S., de Montclos H., Laurent F., Cournoyer B..( 2010;). Epidemiology and molecular characterization of a clone of Burkholderia cenocepacia responsible for nosocomial pulmonary tract infections in a French intensive care unit. Diagn Microbiol Infect Dis66:29–40 [CrossRef][PubMed]
    [Google Scholar]
  25. Hamad M. A., Skeldon A. M., Valvano M. A..( 2010;). Construction of aminoglycoside-sensitive Burkholderia cenocepacia strains for use in studies of intracellular bacteria with the gentamicin protection assay. Appl Environ Microbiol76:3170–3176 [CrossRef][PubMed]
    [Google Scholar]
  26. Hanulik V., Webber M. A., Chroma M., Uvizl R., Holy O., Whitehead R. N., Baugh S., Matouskova I., Kolar M..( 2013;). An outbreak of Burkholderia multivorans beyond cystic fibrosis patients. J Hosp Infect84:248–251 [CrossRef][PubMed]
    [Google Scholar]
  27. Herasimenka Y., Cescutti P., Impallomeni G., Campana S., Taccetti G., Ravenni N., Zanetti F., Rizzo R..( 2007;). Exopolysaccharides produced by clinical strains belonging to the Burkholderia cepacia complex. J Cyst Fibros6:145–152 [CrossRef][PubMed]
    [Google Scholar]
  28. Isles A., Maclusky I., Corey M., Gold R., Prober C., Fleming P., Levison H..( 1984;). Pseudomonas cepacia infection in cystic fibrosis: an emerging problem. J Pediatr104:206–210 [CrossRef][PubMed]
    [Google Scholar]
  29. Jaques A., Daviskas E., Turton J. A., McKay K., Cooper P., Stirling R. G., Robertson C. F., Bye P. T., Lesouëf P. N..& other authors ( 2008;). Inhaled mannitol improves lung function in cystic fibrosis. Chest133:1388–1396 [CrossRef][PubMed]
    [Google Scholar]
  30. Jones A. M., Dodd M. E., Govan J. R., Barcus V., Doherty C. J., Morris J., Webb A. K..( 2004;). Burkholderia cenocepacia and Burkholderia multivorans: influence on survival in cystic fibrosis. Thorax59:948–951 [CrossRef][PubMed]
    [Google Scholar]
  31. Kalish L. A., Waltz D. A., Dovey M., Potter-Bynoe G., McAdam A. J., LiPuma J. J., Gerard C., Goldmann D..( 2006;). Impact of Burkholderia dolosa on lung function and survival in cystic fibrosis. Am J Respir Crit Care Med173:421–425 [CrossRef][PubMed]
    [Google Scholar]
  32. Keith K. E., Valvano M. A..( 2007;). Characterization of SodC, a periplasmic superoxide dismutase from Burkholderia cenocepacia.. Infect Immun75:2451–2460 [CrossRef][PubMed]
    [Google Scholar]
  33. Keith K. E., Killip L., He P., Moran G. R., Valvano M. A..( 2007;). Burkholderia cenocepacia C5424 produces a pigment with antioxidant properties using a homogentisate intermediate. J Bacteriol189:9057–9065 [CrossRef][PubMed]
    [Google Scholar]
  34. Korbsrisate S., Tomaras A. P., Damnin S., Ckumdee J., Srinon V., Lengwehasatit I., Vasil M. L., Suparak S..( 2007;). Characterization of two distinct phospholipase C enzymes from Burkholderia pseudomallei.. Microbiology153:1907–1915 [CrossRef][PubMed]
    [Google Scholar]
  35. Law R. J., Hamlin J. N., Sivro A., McCorrister S. J., Cardama G. A., Cardona S. T..( 2008;). A functional phenylacetic acid catabolic pathway is required for full pathogenicity of Burkholderia cenocepacia in the Caenorhabditis elegans host model. J Bacteriol190:7209–7218 [CrossRef][PubMed]
    [Google Scholar]
  36. Lefebre M., Valvano M..( 2001;). In vitro resistance of Burkholderia cepacia complex isolates to reactive oxygen species in relation to catalase and superoxide dismutase production. Microbiology147:97–109[PubMed]
    [Google Scholar]
  37. Lewenza S., Conway B., Greenberg E. P., Sokol P. A..( 1999;). Quorum sensing in Burkholderia cepacia: identification of the LuxRI homologs CepRI. J Bacteriol181:748–756[PubMed]
    [Google Scholar]
  38. Liou T. G., Adler F. R., Fitzsimmons S. C., Cahill B. C., Hibbs J. R., Marshall B. C..( 2001;). Predictive 5-year survivorship model of cystic fibrosis. Am J Epidemiol153:345–352 [CrossRef][PubMed]
    [Google Scholar]
  39. LiPuma J. J..( 2010;). The changing microbial epidemiology in cystic fibrosis. Clin Microbiol Rev23:299–323 [CrossRef][PubMed]
    [Google Scholar]
  40. Mahenthiralingam E., Coenye T., Chung J. W., Speert D. P., Govan J. R., Taylor P., Vandamme P..( 2000;). Diagnostically and experimentally useful panel of strains from the Burkholderia cepacia complex. J Clin Microbiol38:910–913[PubMed]
    [Google Scholar]
  41. Mann T., Ben-David D., Zlotkin A., Shachar D., Keller N., Toren A., Nagler A., Smollan G., Barzilai A., Rahav G..( 2010;). An outbreak of Burkholderia cenocepacia bacteremia in immunocompromised oncology patients. Infection38:187–194 [CrossRef][PubMed]
    [Google Scholar]
  42. Messiaen A. S., Nelis H., Coenye T..( 2013;). Investigating the role of matrix components in protection of Burkholderia cepacia complex biofilms against tobramycin. J Cyst Fibros [CrossRef][PubMed]
    [Google Scholar]
  43. Middleton P. G., Kidd T. J., Williams B..( 2005;). Combination aerosol therapy to treat Burkholderia cepacia complex. Eur Respir J26:305–308 [CrossRef][PubMed]
    [Google Scholar]
  44. Moore R. A., DeShazer D., Reckseidler S., Weissman A., Woods D. E..( 1999;). Efflux-mediated aminoglycoside and macrolide resistance in Burkholderia pseudomallei.. Antimicrob Agents Chemother43:465–470[PubMed]
    [Google Scholar]
  45. Moreira L. M., Videira P. A., Sousa S. A., Leitão J. H., Cunha M. V., Sá-Correia I..( 2003;). Identification and physical organization of the gene cluster involved in the biosynthesis of Burkholderia cepacia complex exopolysaccharide. Biochem Biophys Res Commun312:323–333 [CrossRef][PubMed]
    [Google Scholar]
  46. Palmer K. L., Aye L. M., Whiteley M..( 2007;). Nutritional cues control Pseudomonas aeruginosa multicellular behavior in cystic fibrosis sputum. J Bacteriol189:8079–8087 [CrossRef][PubMed]
    [Google Scholar]
  47. Rose H., Baldwin A., Dowson C. G., Mahenthiralingam E..( 2009;). Biocide susceptibility of the Burkholderia cepacia complex. J Antimicrob Chemother63:502–510 [CrossRef][PubMed]
    [Google Scholar]
  48. Sage A., Linker A., Evans L. R., Lessie T. G..( 1990;). Hexose phosphate metabolism and exopolysaccharide formation in Pseudomonas cepacia.. Curr Microbiol20:191–198 [CrossRef]
    [Google Scholar]
  49. Sass A., Marchbank A., Tullis E., LiPuma J. J., Mahenthiralingam E..( 2011;). Spontaneous and evolutionary changes in the antibiotic resistance of Burkholderia cenocepacia observed by global gene expression analysis. BMC Genomics12:373. [CrossRef][PubMed]
    [Google Scholar]
  50. Sass A. M., Schmerk C., Agnoli K., Norville P. J., Eberl L., Valvano M. A., Mahenthiralingam E..( 2013;). The unexpected discovery of a novel low-oxygen-activated locus for the anoxic persistence of Burkholderia cenocepacia.. ISME J7:1568–1581 [CrossRef][PubMed]
    [Google Scholar]
  51. Schmerk C. L., Valvano M. A..( 2013;). Burkholderia multivorans survival and trafficking within macrophages. J Med Microbiol62:173–184 [CrossRef][PubMed]
    [Google Scholar]
  52. Shafiq I., Carroll M. P., Nightingale J. A., Daniels T. V..( 2011;). Cepacia syndrome in a cystic fibrosis patient colonised with Burkholderia multivorans.. BMJ Case Rep2011: [CrossRef][PubMed]
    [Google Scholar]
  53. Sokol P. A., Darling P., Woods D. E., Mahenthiralingam E., Kooi C..( 1999;). Role of ornibactin biosynthesis in the virulence of Burkholderia cepacia: characterization of pvdA, the gene encoding L-ornithine N(5)-oxygenase. Infect Immun67:4443–4455[PubMed]
    [Google Scholar]
  54. Sokol P. A., Darling P., Lewenza S., Corbett C. R., Kooi C. D..( 2000;). Identification of a siderophore receptor required for ferric ornibactin uptake in Burkholderia cepacia.. Infect Immun68:6554–6560 [CrossRef][PubMed]
    [Google Scholar]
  55. Subsin B., Chambers C. E., Visser M. B., Sokol P. A..( 2007;). Identification of genes regulated by the cepIR quorum-sensing system in Burkholderia cenocepacia by high-throughput screening of a random promoter library. J Bacteriol189:968–979 [CrossRef][PubMed]
    [Google Scholar]
  56. Tatusov R. L., Galperin M. Y., Natale D. A., Koonin E. V..( 2000;). The COG database: a tool for genome-scale analysis of protein functions and evolution. Nucleic Acids Res28:33–36 [CrossRef][PubMed]
    [Google Scholar]
  57. Teper A., Jaques A., Charlton B..( 2011;). Inhaled mannitol in patients with cystic fibrosis: A randomised open-label dose response trial. J Cyst Fibros10:1–8 [CrossRef][PubMed]
    [Google Scholar]
  58. Tomich M., Herfst C. A., Golden J. W., Mohr C. D..( 2002;). Role of flagella in host cell invasion by Burkholderia cepacia.. Infect Immun70:1799–1806 [CrossRef][PubMed]
    [Google Scholar]
  59. Tseng B. S., Zhang W., Harrison J. J., Quach T. P., Song J. L., Penterman J., Singh P. K., Chopp D. L., Packman A. I., Parsek M. R..( 2013;). The extracellular matrix protects Pseudomonas aeruginosa biofilms by limiting the penetration of tobramycin. Environ Microbiol15:2865–2878 [CrossRef][PubMed]
    [Google Scholar]
  60. Tunpiboonsak S., Mongkolrob R., Kitudomsub K., Thanwatanaying P., Kiettipirodom W., Tungboontina Y., Tungpradabkul S..( 2010;). Role of a Burkholderia pseudomallei polyphosphate kinase in an oxidative stress response, motilities, and biofilm formation. J Microbiol48:63–70 [CrossRef][PubMed]
    [Google Scholar]
  61. Urban T. A., Griffith A., Torok A. M., Smolkin M. E., Burns J. L., Goldberg J. B..( 2004;). Contribution of Burkholderia cenocepacia flagella to infectivity and inflammation. Infect Immun72:5126–5134 [CrossRef][PubMed]
    [Google Scholar]
  62. Vial L., Chapalain A., Groleau M. C., Déziel E..( 2011;). The various lifestyles of the Burkholderia cepacia complex species: a tribute to adaptation. Environ Microbiol13:1–12 [CrossRef][PubMed]
    [Google Scholar]
  63. Whiteford M. L., Wilkinson J. D., McColl J. H., Conlon F. M., Michie J. R., Evans T. J., Paton J. Y..( 1995;). Outcome of Burkholderia (Pseudomonas) cepacia colonisation in children with cystic fibrosis following a hospital outbreak. Thorax50:1194–1198 [CrossRef][PubMed]
    [Google Scholar]
  64. Winsor G. L., Khaira B., Van Rossum T., Lo R., Whiteside M. D., Brinkman F. S..( 2008;). The Burkholderia Genome Database: facilitating flexible queries and comparative analyses. Bioinformatics24:2803–2804 [CrossRef][PubMed]
    [Google Scholar]
  65. Woods C. W., Bressler A. M., LiPuma J. J., Alexander B. D., Clements D. A., Weber D. J., Moore C. M., Reller L. B., Kaye K. S..( 2004;). Virulence associated with outbreak-related strains of Burkholderia cepacia complex among a cohort of patients with bacteremia. Clin Infect Dis38:1243–1250 [CrossRef][PubMed]
    [Google Scholar]
  66. Zahariadis G., Levy M. H., Burns J. L..( 2003;). Cepacia-like syndrome caused by Burkholderia multivorans.. Can J Infect Dis Med Microbiol14:123–125[PubMed]
    [Google Scholar]
  67. Zlosnik J. E., Costa P. S., Brant R., Mori P. Y., Hird T. J., Fraenkel M. C., Wilcox P. G., Davidson A. G., Speert D. P..( 2011;). Mucoid and nonmucoid Burkholderia cepacia complex bacteria in cystic fibrosis infections. Am J Respir Crit Care Med183:67–72 [CrossRef][PubMed]
    [Google Scholar]
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