Proteomic, microarray, and signature-tagged mutagenesis analyses of anaerobic Pseudomonas aeruginosa at pH 6.5, likely representing chronic, late-stage cystic fibrosis airway conditions

Mark D. Platt, Michael J. Schurr, Karin Sauer, Gustavo Vazquez, Irena Kukavica-Ibrulj, Eric Potvin, Roger C. Levesque, Amber Fedynak, Fiona S.L. Brinkman, Jill Schurr, Sung Hei Hwang, Gee W. Lau, Patrick A. Limbach, John J. Rowe, Michael A. Lieberman, Nicolas Barraud, Jeremy Webb, Staffan Kjelleberg, Donald F. Hunt, Daniel J. Hassett

Research output: Contribution to journalArticle

Abstract

Patients suffering from cystic fibrosis (CF) commonly harbor the important pathogen Pseudomonas aeruginosa in their airways. During chronic late-stage CF, P. aeruginosa is known to grow under reduced oxygen tension and is even capable of respiring anaerobically within the thickened airway mucus, at a pH of ∼6.5. Therefore, proteins involved in anaerobic metabolism represent potentially important targets for therapeutic intervention. In this study, the clinically relevant "anaerobiome" or "proteogenome" of P. aeruginosa was assessed. First, two different proteomic approaches were used to identify proteins differentially expressed under anaerobic versus aerobic conditions. Microarray studies were also performed, and in general, the anaerobic transcriptome was in agreement with the proteomic results. However, we found that a major portion of the most upregulated genes in the presence of NO3- and NO2- are those encoding Pf1 bacteriophage. With anaerobic NO2-, the most downregulated genes are those involved postglycolytically and include many tricarboxylic acid cycle genes and those involved in the electron transport chain, especially those encoding the NADH dehydrogenase I complex. Finally, a signature-tagged mutagenesis library of P. aeruginosa was constructed to further screen genes required for both NO3- and NO 2- respiration. In addition to genes anticipated to play important roles in the anaerobiome (anr, dnr, nar, nir, and nuo), the cysG and dksA genes were found to be required for both anaerobic NO3 - and NO2- respiration. This study represents a major step in unraveling the molecular machinery involved in anaerobic NO 3- and NO2- respiration and offers clues as to how we might disrupt such pathways in P. aeruginosa to limit the growth of this important CF pathogen when it is either limited or completely restricted in its oxygen supply.

Original languageEnglish (US)
Pages (from-to)2739-2758
Number of pages20
JournalJournal of bacteriology
Volume190
Issue number8
DOIs
StatePublished - Apr 1 2008
Externally publishedYes

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Cystic Fibrosis
Mutagenesis
Proteomics
Pseudomonas aeruginosa
Genes
Respiration
Bacteriophage Pf1
Oxygen
Anaerobiosis
Electron Transport Complex I
Citric Acid Cycle
Mucus
Electron Transport
Transcriptome
Libraries
Proteins
Down-Regulation
Growth

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Proteomic, microarray, and signature-tagged mutagenesis analyses of anaerobic Pseudomonas aeruginosa at pH 6.5, likely representing chronic, late-stage cystic fibrosis airway conditions. / Platt, Mark D.; Schurr, Michael J.; Sauer, Karin; Vazquez, Gustavo; Kukavica-Ibrulj, Irena; Potvin, Eric; Levesque, Roger C.; Fedynak, Amber; Brinkman, Fiona S.L.; Schurr, Jill; Hwang, Sung Hei; Lau, Gee W.; Limbach, Patrick A.; Rowe, John J.; Lieberman, Michael A.; Barraud, Nicolas; Webb, Jeremy; Kjelleberg, Staffan; Hunt, Donald F.; Hassett, Daniel J.

In: Journal of bacteriology, Vol. 190, No. 8, 01.04.2008, p. 2739-2758.

Research output: Contribution to journalArticle

Platt, MD, Schurr, MJ, Sauer, K, Vazquez, G, Kukavica-Ibrulj, I, Potvin, E, Levesque, RC, Fedynak, A, Brinkman, FSL, Schurr, J, Hwang, SH, Lau, GW, Limbach, PA, Rowe, JJ, Lieberman, MA, Barraud, N, Webb, J, Kjelleberg, S, Hunt, DF & Hassett, DJ 2008, 'Proteomic, microarray, and signature-tagged mutagenesis analyses of anaerobic Pseudomonas aeruginosa at pH 6.5, likely representing chronic, late-stage cystic fibrosis airway conditions', Journal of bacteriology, vol. 190, no. 8, pp. 2739-2758. https://doi.org/10.1128/JB.01683-07
Platt, Mark D. ; Schurr, Michael J. ; Sauer, Karin ; Vazquez, Gustavo ; Kukavica-Ibrulj, Irena ; Potvin, Eric ; Levesque, Roger C. ; Fedynak, Amber ; Brinkman, Fiona S.L. ; Schurr, Jill ; Hwang, Sung Hei ; Lau, Gee W. ; Limbach, Patrick A. ; Rowe, John J. ; Lieberman, Michael A. ; Barraud, Nicolas ; Webb, Jeremy ; Kjelleberg, Staffan ; Hunt, Donald F. ; Hassett, Daniel J. / Proteomic, microarray, and signature-tagged mutagenesis analyses of anaerobic Pseudomonas aeruginosa at pH 6.5, likely representing chronic, late-stage cystic fibrosis airway conditions. In: Journal of bacteriology. 2008 ; Vol. 190, No. 8. pp. 2739-2758.
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AU - Sauer, Karin

AU - Vazquez, Gustavo

AU - Kukavica-Ibrulj, Irena

AU - Potvin, Eric

AU - Levesque, Roger C.

AU - Fedynak, Amber

AU - Brinkman, Fiona S.L.

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AU - Hwang, Sung Hei

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AU - Lieberman, Michael A.

AU - Barraud, Nicolas

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AU - Hunt, Donald F.

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