Nitrate transport and its regulation by O2 in Pseudomonas aeruginosa

Hernandez, Dennis; Dias, Fiona M.; Rowe, John J.

doi:10.1016/0003-9861(91)90022-b

Cited by 23 publications

(19 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It has been shown that ANR is a global regulator for anaerobiosis, regulating the anaerobic induction of the genes required for denitrification (Arai et al, 1995;Ye et al, 1995), arginine deiminase pathway, and cyanide production (Hassett and Zak, 2005;Zimmermann et al, 1991). Post-translational inhibition by oxygen on denitrification enzymes and nitrate transport in P. aeruginosa has also been reported Rowe, 1987, 1988;Hernandez et al, 1991). Nonetheless, Arai et al (1995) have demonstrated that ANR is not necessarily required for denitrification by P. aeruginosa when another CRP (catabolite regulator protein)/FNRrelated transcriptional regulator, DNR, is compulsorily expressed.…”

Section: Introductionmentioning

confidence: 92%

Competition between oxygen and nitrate respirations in continuous culture of Pseudomonas aeruginosa performing aerobic denitrification

Chen

Xia

2006

Biotech & Bioengineering

View full text Add to dashboard Cite

Continuous culture of P. aeruginosa was conducted with nitrate-containing media under the dilution rates (D) of 0.026, 0.06, and 0.13/h and the dissolved oxygen concentrations (DO) of 0-2.2 mg/L. The bacterium performed simultaneous O(2) and nitrate respiration in all of the systems studied. For each D, the (apparent) cell yield from glucose (Y(X/S)) was lower at zero DO, but did not change substantially with non-zero DO. In non-zero DO systems, Y(X/S) increased with increasing D, and when fit with a model considering cell death, gave the following parameters: maximum cell yield Y(X/S) (m) = 0.49, maintenance coefficient M(S) = 0.029 (/h), and cell decay constant k(d) = 0.014/h. The same model failed to describe the behaviors of zero-DO systems, where neither glucose nor nitrate was limiting and the limiting factor(s) remained unknown. The cell yield from accepted electron (Y(X/e)) was however relatively constant in all systems, and the energy yield per electron accepted via denitrification was estimated at approximately 69% of that via O(2) respiration. A closer examination revealed that increasing DO enhanced O(2) respiration only at extremely low DO ( <0.05 mg/L), beyond which the increasing DO only slightly increased its weak inhibition on denitrification. While O(2) was the preferred electron acceptor, the fraction of electrons accepted via denitrification increased with increasing D.

show abstract

Section: Introductionmentioning

confidence: 92%

Competition between oxygen and nitrate respirations in continuous culture of Pseudomonas aeruginosa performing aerobic denitrification

Chen

Xia

2006

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…Upon uptake, Mo is introduced into a complex molybdopterin molecule to generate the molybdenum cofactor (MoCo), which may then be further modified prior to insertion into MoCo-dependent proteins (13). Such proteins include dimethyl sulfoxide (DMSO) reductase, xanthine oxidase, and sulfite oxidase, all of which have broad roles in nitrogen, carbon, and sulfur metabolism (13,14). Acquisition of Mo by prokaryotes occurs primarily via a high-affinity ATP-binding cassette (ABC) permease, ModABC.…”

mentioning

confidence: 99%

Acquisition and Role of Molybdate in Pseudomonas aeruginosa

Pederick

Eijkelkamp

Ween

et al. 2014

Appl Environ Microbiol

View full text Add to dashboard Cite

In microaerophilic or anaerobic environments, Pseudomonas aeruginosa utilizes nitrate reduction for energy production, a process dependent on the availability of the oxyanionic form of molybdenum, molybdate (MoO 4 2؊ ). Here, we show that molybdate acquisition in P. aeruginosa occurs via a high-affinity ATP-binding cassette permease (ModABC). ModA is a cluster D-III solute binding protein capable of interacting with molybdate or tungstate oxyanions. Deletion of the modA gene reduces cellular molybdate concentrations and results in inhibition of anaerobic growth and nitrate reduction. Further, we show that conditions that permit nitrate reduction also cause inhibition of biofilm formation and an alteration in fatty acid composition of P. aeruginosa. Collectively, these data highlight the importance of molybdate for anaerobic growth of P. aeruginosa and reveal novel consequences of nitrate reduction on biofilm formation and cell membrane composition.

show abstract

“…Parallel studies of P. aeruginosa have resulted in the characterization of a unique nar operon (24, 27) regulated by the proteins Anr and Dnr (40) as well as narX and narL (24). However, a narQ homologue has not been identified (30,34).Posttranslationally, oxygen also has the capacity to inhibit denitrification immediately at the level of nitrate uptake and nitrite excretion (10,11,33) as well as through the diversion of electron flow to oxygen in E. coli and in Paracoccus denitrificans (4, 33). Despite these studies, an experimental method for the measurement of posttranslational regulation by oxygen has been lacking.…”

mentioning

confidence: 99%

“…Posttranslationally, oxygen also has the capacity to inhibit denitrification immediately at the level of nitrate uptake and nitrite excretion (10,11,33) as well as through the diversion of electron flow to oxygen in E. coli and in Paracoccus denitrificans (4, 33). Despite these studies, an experimental method for the measurement of posttranslational regulation by oxygen has been lacking.…”

mentioning

confidence: 99%

“…Although similar nitrate reductase activities were observed in aerobic JVC and anaerobic JVC with pREP (Table 3), the levels of nitrite excretion were significantly lower under aerobic conditions. The P LAC insertion enabled JVC to aerobically transcribe nar and produce functional wild-type levels of respiratory nitrate reductase activity under aerobic conditions, thus clearly demonstrating the physiological significance of posttranslational regulation by oxygen at the level of nitrate transport and/or diversion of electron flow (4,10,11).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Artificial Control of Nitrate Respiration through the lac Promoter Permits the Assessment of Oxygen-Mediated Posttranslational Regulation of the nar Operon in Pseudomonas aeruginosa

2007

Self Cite

View full text Add to dashboard Cite

In this study, oxygen and nitrate regulation of transcription and subsequent protein expression of the unique narK1K2GHJI respiratory operon of Pseudomonas aeruginosa were investigated. Under the control of P LAC , P. aeruginosa was able to transcribe nar and subsequently express methyl viologen-linked nitrate reductase activity under aerobic conditions without nitrate. Modulation of P LAC through the LacI repressor enabled us to assess both transcriptional and posttranslational regulation by oxygen during physiological whole-cell nitrate reduction.Pseudomonas aeruginosa is a ubiquitous gram-negative bacterium capable of growth and/or survival anaerobically through arginine catabolism (36), pyruvate fermentation (6), or denitrification in the presence of nitrogen oxides (37). The latter process allows this organism to persist in soil as part of the global nitrogen cycle. Additionally, denitrification has been implicated in infections by this opportunistic pathogen in the airways of cystic fibrosis patients (9,18,35).During anaerobic growth of Escherichia coli, the Fnr protein is responsible for activation of the synthesis of anaerobic respiratory enzymes such as nitrate reductase (28,31). In addition, the presence of external nitrate induces the transcription of the nitrate reductase operon through the dual twocomponent regulatory systems of narX-narL (31, 32) and narQnarP (3,21,22). Parallel studies of P. aeruginosa have resulted in the characterization of a unique nar operon (24, 27) regulated by the proteins Anr and Dnr (40) as well as narX and narL (24). However, a narQ homologue has not been identified (30,34).Posttranslationally, oxygen also has the capacity to inhibit denitrification immediately at the level of nitrate uptake and nitrite excretion (10,11,33) as well as through the diversion of electron flow to oxygen in E. coli and in Paracoccus denitrificans (4, 33). Despite these studies, an experimental method for the measurement of posttranslational regulation by oxygen has been lacking.In the present study, the effects of oxygen and nitrate on the expression of the narK1K2GHJI operon (27) were examined during aerobic or anaerobic growth with and without nitrate. In addition, a P LAC element was inserted upstream of the respiratory nitrate reductase genes (narK2GHJI) of P. aeruginosa to overcome transcriptional regulation of the nar operon by oxygen and nitrate. The levels of transcription, respiratory nitrate reductase activity, and whole-cell physiological reduction of nitrate to nitrite were measured under both aerobic and anaerobic conditions, thus allowing quantitative assessment of posttranslational regulation by oxygen.The bacterial strains and plasmids used in this study are listed in Table 1. All bacteria were grown at 37°C from singlecolony isolates or overnight cultures in Luria-Bertani (LB) broth (Fisher Scientific, Pittsburgh, PA). The medium was supplemented with 1% (wt/vol) KNO 3 (LB-NO 3 ) when indicated. Aerobic cultures were set up as 50-ml volumes of LB or LB-NO 3 in a 500-ml Erlenm...

show abstract

Nitrate transport and its regulation by O2 in Pseudomonas aeruginosa

Cited by 23 publications

References 38 publications

Competition between oxygen and nitrate respirations in continuous culture of Pseudomonas aeruginosa performing aerobic denitrification

Competition between oxygen and nitrate respirations in continuous culture of Pseudomonas aeruginosa performing aerobic denitrification

Acquisition and Role of Molybdate in Pseudomonas aeruginosa

Artificial Control of Nitrate Respiration through the lac Promoter Permits the Assessment of Oxygen-Mediated Posttranslational Regulation of the nar Operon in Pseudomonas aeruginosa

Contact Info

Product

Resources

About