Phenazines regulate Napdependent denitrification in Pseudomonas aeruginosa biofilms

Yu Cheng Lin, Matthew D. Sekedat, William Cole Cornell, Gustavo M. Silva, Chinweike Okegbe, Alexa Price-Whelan, Christine Vogel, Lars E.P. Dietrich

Research output: Contribution to journalArticle

Abstract

Microbes in biofilms face the challenge of substrate limitation. In particular, oxygen often becomes limited for cells in Pseudomonas aeruginosa biofilms growing in the laboratory or during host colonization. Previously we found that phenazines, antibiotics produced by P. aeruginosa, balance the intracellular redox state of cells in biofilms. Here, we show that genes involved in denitrification are induced in phenazine-null (Δphz) mutant biofilms grown under an aerobic atmosphere, even in the absence of nitrate. This finding suggests that resident cells employ a bet-hedging strategy to anticipate the potential availability of nitrate and counterbalance their highly reduced redox state. Consistent with our previous characterization of aerobically grown colonies supplemented with nitrate, we found that the pathway that is induced in Δphz mutant colonies combines the nitrate reductase activity of the periplasmic enzyme Nap with the downstream reduction of nitrite to nitrogen gas catalyzed by the enzymes Nir, Nor, and Nos. This regulatory relationship differs from the denitrification pathway that functions under anaerobic growth, with nitrate as the terminal electron acceptor, which depends on the membrane-associated nitrate reductase Nar. We identified the sequences in the promoter regions of the nap and nir operons that are required for the effects of phenazines on expression. We also show that specific phenazines have differential effects on nap gene expression. Finally, we provide evidence that individual steps of the denitrification pathway are catalyzed at different depths within aerobically grown biofilms, suggesting metabolic cross-feeding between community subpopulations.

Original languageEnglish (US)
Article numbere00031-18
JournalJournal of bacteriology
Volume200
Issue number9
DOIs
StatePublished - May 1 2018

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Keywords

  • Anr
  • Biofilm physiology
  • Denitrification
  • Nap
  • Nir
  • Nitrate reductase
  • Nitrite reductase
  • RpoS

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Lin, Y. C., Sekedat, M. D., Cornell, W. C., Silva, G. M., Okegbe, C., Price-Whelan, A., Vogel, C., & Dietrich, L. E. P. (2018). Phenazines regulate Napdependent denitrification in Pseudomonas aeruginosa biofilms. Journal of bacteriology, 200(9), [e00031-18]. https://doi.org/10.1128/JB.00031-18