1887

Abstract

The opportunistic bacterial pathogen causes acute and chronic infections that are notoriously difficult to treat. In people with cystic fibrosis, can cause lifelong lung infections, and isolation of mucoid , resulting from the overproduction of alginate, is associated with chronic infection. The histone-like protein AlgP has previously been implicated in the control of alginate gene expression in mucoid strains, but this regulation is unclear. To explore AlgP in further detail, we deleted in mucoid strains and demonstrated that the deletion of did not result in a nonmucoid phenotype or a decrease in alginate production. We showed that the promoter is expressed by both the nonmucoid strain PAO1 and the isogenic mucoid strain PDO300, suggesting that there may be genes that are differentially regulated between these strains. In support of this, using RNA sequencing, we identified a small AlgP regulon that has no significant overlap between PAO1 and PDO300 and established that alginate genes were not differentially regulated by the deletion of . Of note, we found that deleting in PAO1 increased expression of the nitric oxide operon and the nitrous oxide reductase genes and subsequently promoted growth of PAO1 under anaerobic conditions. Altogether, we have defined a narrow regulon of genes controlled by AlgP and provided evidence that alginate production is not greatly affected by AlgP, countering the long-standing premise in the field.

Funding
This study was supported by the:
  • National Institutes of Health (US) (Award R35 GM119426)
    • Principle Award Recipient: William M. Wuest
  • Cystic Fibrosis Foundation (Award WHITEL16G0)
    • Principle Award Recipient: Marvin Whiteley
  • Cystic Fibrosis Foundation (US) (Award WHITEL19P0)
    • Principle Award Recipient: Marvin Whiteley
  • National Institutes of Health (US) (Award R56 HL142857)
    • Principle Award Recipient: Marvin Whiteley
  • National Institutes of Health (US) (Award R01 GM116547)
    • Principle Award Recipient: Marvin Whiteley
  • Cystic Fibrosis Foundation (Award GOLDBE16G0)
    • Principle Award Recipient: Joanna B Goldberg
  • Cystic Fibrosis Foundation (US) (Award MCCART15R0)
    • Principle Award Recipient: Ashley R. Cross
  • National Institutes of Health (Award F31 AI136310)
    • Principle Award Recipient: Ashley R. Cross
  • National Institutes of Health (US) (Award R21 AI122192)
    • Principle Award Recipient: Joanna B Goldberg
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2020-07-07
2021-10-28
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References

  1. Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P et al. Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 2000; 406:959–964 [View Article]
    [Google Scholar]
  2. Bauernfeind A, Hörl G, Jungwirth R, Petermüller C, Przyklenk B et al. Qualitative and quantitative microbiological analysis of sputa of 102 patients with cystic fibrosis. Infection 1987; 15:270–277 [View Article]
    [Google Scholar]
  3. Mearns MB, Hunt GH, Rushworth R. Bacterial flora of respiratory tract in patients with cystic fibrosis, 1950-71. Arch Dis Child 1972; 47:902–907 [View Article][PubMed]
    [Google Scholar]
  4. Cystic Fibrosis Foundation Cystic fibrosis patient registry annual data report; 2016
  5. Quinton PM. Cystic fibrosis: impaired bicarbonate secretion and mucoviscidosis. The Lancet 2008; 372:415–417 [View Article]
    [Google Scholar]
  6. Quinton PM, Bijman J. Higher bioelectric potentials due to decreased chloride absorption in the sweat glands of patients with cystic fibrosis. New England Journal of Medicine 1983; 308:1185–1189 [View Article]
    [Google Scholar]
  7. Rommens J, Kerem B, Alon N, Rozmahel R et al. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 1989; 245:1066–1073 [View Article]
    [Google Scholar]
  8. Kreda SM, Davis CW, Rose MC. Cftr, mucins, and mucus obstruction in cystic fibrosis. Cold Spring Harb Perspect Med 2012; 2:a009589 [View Article]
    [Google Scholar]
  9. Boucher JC, Yu H, Mudd MH, Deretic V. Mucoid Pseudomonas aeruginosa in cystic fibrosis: characterization of muc mutations in clinical isolates and analysis of clearance in a mouse model of respiratory infection. Infect Immun 1997; 65:3838–3846 [View Article]
    [Google Scholar]
  10. Doggett R, Harrison GM, RE CJ. Mucoid Pseudomonas aeruginosaqRUGINOSA in patients with chronic illnesses. The Lancet 1971; 297:236–237 [View Article]
    [Google Scholar]
  11. Mathee K, Ciofu O, Sternberg C, Lindum PW, Campbell JIA et al. Mucoid conversion of Pseudomonas aeruginos by hydrogen peroxide: a mechanism for virulence activation in the cystic fibrosis lung. Microbiology 1999; 145:1349–1357 [View Article]
    [Google Scholar]
  12. Martin DW, Schurr MJ, Mudd MH, Govan JR, Holloway BW et al. Mechanism of conversion to mucoidy in Pseudomonas aeruginosa infecting cystic fibrosis patients. Proc Natl Acad Sci U S A 1993; 90:8377–8381 [View Article]
    [Google Scholar]
  13. Candido Cacador N. Paulino da Costa Capizzani C, Gomes Monteiro Marin Torres LA, Galetti R, Ciofu O, da Costa Darini AL, et al. Adaptation of Pseudomonas aeruginosa to the chronic phenotype by mutations in the algTmucABD operon in isolates from Brazilian cystic fibrosis patients. PLoS One 2018; 13:e0208013
    [Google Scholar]
  14. Lam J, Chan R, Lam K, Costerton JW. Production of mucoid microcolonies by Pseudomonas aeruginosa within infected lungs in cystic fibrosis. Infect Immun 1980; 28:546–556
    [Google Scholar]
  15. Malhotra S, Hayes D, Wozniak DJ. Mucoid Pseudomonas aeruginosa and regional inflammation in the cystic fibrosis lung. J Cyst Fibros 2019; 18:796–803 [View Article]
    [Google Scholar]
  16. Govan JR, Deretic V. Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia . Microbiol Rev 1996; 60:539–574 [View Article]
    [Google Scholar]
  17. Mathee K, McPherson CJ, Ohman DE. Posttranslational control of the algT (algU)-encoded sigma22 for expression of the alginate regulon in Pseudomonas aeruginosa and localization of its antagonist proteins MucA and MucB (AlgN). J Bacteriol 1997; 179:3711–3720 [View Article]
    [Google Scholar]
  18. Baynham PJ, Wozniak DJ. Identification and characterization of AlgZ, an AlgT-dependent DNA-binding protein required for Pseudomonas aeruginosa algD transcription. Mol Microbiol 1996; 22:97–108 [View Article]
    [Google Scholar]
  19. Wozniak DJ, Ohman DE. Transcriptional analysis of the Pseudomonas aeruginosa genes algR, algB, and algD reveals a hierarchy of alginate gene expression which is modulated by algT . J Bacteriol 1994; 176:6007–6014 [View Article]
    [Google Scholar]
  20. Wozniak DJ, Ohman DE. Pseudomonas aeruginosa algB, a two-component response regulator of the NtrC family, is required for algD transcription. J Bacteriol 1991; 173:1406–1413 [View Article]
    [Google Scholar]
  21. Goldberg JB, Gorman WL, Flynn JL, Ohman DE. A mutation in algN permits trans activation of alginate production by algT in Pseudomonas species. J Bacteriol 1993; 175:1303–1308 [View Article]
    [Google Scholar]
  22. Wood LF, Leech AJ, Ohman DE. Cell wall-inhibitory antibiotics activate the alginate biosynthesis operon in Pseudomonas aeruginosa: roles of sigma (AlgT) and the AlgW and Prc proteases. Mol Microbiol 2006; 62:412–426 [View Article]
    [Google Scholar]
  23. Diggle SP, Winzer K, Lazdunski Andrée, Williams P, Cámara M. Advancing the quorum in Pseudomonas aeruginosa: MvaT and the regulation of N-acylhomoserine lactone production and virulence gene expression. J Bacteriol 2002; 184:2576–2586 [View Article]
    [Google Scholar]
  24. Vallet I, Diggle SP, Stacey RE, Cámara M, Ventre I et al. Biofilm formation in Pseudomonas aeruginosa: fimbrial cup gene clusters are controlled by the transcriptional regulator MvaT. J Bacteriol 2004; 186:2880–2890 [View Article]
    [Google Scholar]
  25. Toussaint B, Delicattree I, Vignais PM. Pseudomonas aeruginosa contains an IHF-like protein that binds to the algD promoter. Biochem Biophys Res Commun 1993; 196:416–421 [View Article]
    [Google Scholar]
  26. Konyecsni WM, Deretic V. DNA sequence and expression analysis of algP and algQ, components of the multigene system transcriptionally regulating mucoidy in Pseudomonas aeruginosa: algP contains multiple direct repeats . J Bacteriol 1990; 172:2511–2520 [View Article]
    [Google Scholar]
  27. Kato J, Misra TK, Chakrabarty AM. AlgR3, a protein resembling eukaryotic histone H1, regulates alginate synthesis in Pseudomonas aeruginosa . Proc Natl Acad Sci U S A 1990; 87:2887–2891 [View Article]
    [Google Scholar]
  28. Deretic V, Hibler NS, Holt SC. Immunocytochemical analysis of AlgP (HP1), a histonelike element participating in control of mucoidy in Pseudomonas aeruginosa . J Bacteriol 1992; 174:824–831 [View Article]
    [Google Scholar]
  29. Deretic V, Konyecsni WM. A procaryotic regulatory factor with a histone H1-like carboxy-terminal domain: clonal variation of repeats within algP, a gene involved in regulation of mucoidy in Pseudomonas aeruginosa . J Bacteriol 1990; 172:5544–5554 [View Article]
    [Google Scholar]
  30. Pier GB, Matthews WJ, Eardley DD. Immunochemical characterization of the mucoid exopolysaccharide of Pseudomonas aeruginosa . J Infect Dis 1983; 147:494–503 [View Article]
    [Google Scholar]
  31. Wood LF, Ohman DE. Identification of genes in the sigma(22) regulon of Pseudomonas aeruginosa required for cell envelope homeostasis in either the planktonic or the sessile mode of growth. mBio 2012; 3: [View Article]
    [Google Scholar]
  32. Deretic V, Gill JF, Chakrabarty AM. Gene algD coding for GDPmannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa . J Bacteriol 1987; 169:351–358 [View Article]
    [Google Scholar]
  33. Firoved AM, Deretic V. Microarray analysis of global gene expression in mucoid Pseudomonas aeruginosa . J Bacteriol 2003; 185:1071–1081 [View Article]
    [Google Scholar]
  34. Yoon SS, Karabulut AC, Lipscomb JD, Hennigan RF, Lymar SV et al. Two-pronged survival strategy for the major cystic fibrosis pathogen, Pseudomonas aeruginosa, lacking the capacity to degrade nitric oxide during anaerobic respiration. EMBO J 2007; 26:3662–3672 [View Article]
    [Google Scholar]
  35. Tunney MM, Field TR, Moriarty TF, Patrick S, Doering G et al. Detection of anaerobic bacteria in high numbers in sputum from patients with cystic fibrosis. Am J Respir Crit Care Med 2008; 177:995–1001 [View Article]
    [Google Scholar]
  36. Worlitzsch D, Tarran R, Ulrich M, Schwab U, Cekici A et al. Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients. J Clin Invest 2002; 109:317–325 [View Article][PubMed]
    [Google Scholar]
  37. Weinging Z, Cross AR, Crowe-McAuliffe C, Weigert-Munoz A, Csatary EE et al. The natural product elegaphenone potentiates antibiotic effects against Pseudomonas aeruginosa . Angew Chem Int Ed 2019; 58:1–5 .
    [Google Scholar]
  38. Babin BM, Atangcho L, van Eldijk MB, Sweredoski MJ, Moradian A et al. Selective proteomic analysis of Antibiotic-Tolerant cellular subpopulations in Pseudomonas aeruginosa biofilms. mBio 2017; 8: [View Article]
    [Google Scholar]
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