Regulation of expression of the cyanide-insensitive terminal oxidase in Pseudomonas aeruginosa

Abstract

The regulation of the cyanide-insensitive oxidase (CIO) in Pseudomonas aeruginosa, a bacterium that can synthesize HCN, is reported. The expression of a cioA-lacZ transcriptional fusion, CioA protein levels and CIO activity were low in exponential phase but induced about fivefold upon entry into stationary phase. Varying the O 2 transfer coefficient from 11?5 h 21 to 87?4 h 21 had no effect on CIO expression and no correlation was observed between CIO induction and the dissolved O 2 levels in the growth medium. However, a mutant deleted for the O 2 -sensitive transcriptional regulator ANR derepressed CIO expression in an O 2 -sensitive manner, with the highest induction occurring under low-O 2 conditions. Therefore, CIO expression can respond to a signal generated by low O 2 levels, but this response is normally kept in check by ANR repression. ANR may play an important role in preventing overexpression of the CIO in relation to other terminal oxidases. A component present in spent culture medium was able to induce CIO expression. However, experiments with purified N-butanoyl-L-homoserine lactone or N-(3-oxododecanoyl)homoserine lactone ruled out a role for these quorum-sensing molecules in the control of CIO expression. Cyanide was a potent inducer of the CIO at physiologically relevant concentrations and experiments using spent culture medium from a DhcnB mutant, which is unable to synthesize cyanide, showed that cyanide was the inducing factor present in P. aeruginosa spent culture medium. However, the finding that in a DhcnB mutant cioA-lacZ expression was induced normally upon entry into stationary phase indicated that cyanide was not the endogenous inducer of the terminal oxidase. The authors suggest that the failure of O 2 to have an effect on CIO expression in the wild-type can be explained either by the requirement for an additional, stationary-phase-specific inducing signal or by the loss of an exponential-phase-specific repressing signal. INTRODUCTIONPseudomonas aeruginosa is an opportunistic pathogen that causes a variety of nosocomial infections, including pneumonia, urinary tract infections, surgical wound infections, and bloodstream infections (for a review see Deretic, 2000). It causes life-threatening illness in patients with cystic fibrosis. Initially, P. aeruginosa colonizes the airways with other pathogens such as Haemophilus influenzae and Staphylococcus aureus. However, in most of these patients chronic lung disease develops in which the bacterial population consists almost exclusively of P. aeruginosa in the form of biofilms (Govan & Deretic, 1996). P. aeruginosa is a facultative anaerobe that preferentially obtains its energy via aerobic respiration, but it is well adapted to conditions of limited O 2 supply (Palleroni, 1984;Davies et al., 1989). It is capable of anaerobic growth with nitrate as a terminal electron acceptor and in the absence of nitrate it is able to ferment arginine, generating ATP by substrate-level phosphorylation (Palleroni, 1984;Davies et al., 1989;Van der Wauven et ...

Enzymatic Characterization and In Vivo Function of Five Terminal Oxidases in Pseudomonas aeruginosa

“…The H ϩ /O ratio of the entire ETC terminated by CIO was lowest among the ETCs involving the other terminal oxidases, indicating that the utilization of CIO is not efficient for the generation of ATP. CIO is induced by inhibitors of the respiratory chain, cyanide and sodium nitroprusside (SNP), or by copper starvation (16,18,19,21). Thus, CIO acts as a complementary enzyme when the heme-copper oxidases are not functioning or are inhibited.…”

“…Inactivation of the other terminal oxidases by respiratory inhibitors or gene disruption also leads to significant upregulation of the cio genes (15)(16)(17)(18)(19)(20)(21). The upregulation of CIO in the stationary phase is probably because P. aeruginosa produces cyanide, which inhibits heme-copper oxidases, in the stationary phase.…”

“…The upregulation of CIO in the stationary phase is probably because P. aeruginosa produces cyanide, which inhibits heme-copper oxidases, in the stationary phase. The cio operon is positively regulated by RoxSR, which is a two-component regulatory system that may sense respiratory chain electron flow or redox status of the quinone pool, and is negatively regulated by ANR (anaerobic regulator of arginine deiminase and nitrate reductase), which is a low-oxygen-responsive global regulator (17,18). The stationary-phase sigma factor RpoS also activates expression of the cio genes (21).…”

Cell aggregation of Pseudomonas aeruginosa strain PAO1 as an energy-dependent stress response during growth with sodium dodecyl sulfate

“…Both inhibitors also reduced degradation of the respective substrates to the same extent as they inhibited respiration (not shown). To account for the influence of cyanide-insensitive respiration (Cunningham et al 1997), we investigated a cioB mutant which is defective in the cyanide-insensitive cytochrome c oxidase Cio (Cooper et al 2003). KCN inhibited the substrate-dependent oxygen uptake in this mutant completely (Table 1).…”

Adaptation of Aerobically GrowingPseudomonas aeruginosato Copper Starvation

“…By controlling the expression of multiple cytochrome oxidases, P. aeruginosa appears to exploit the best-suited electron transport chain in response to the available oxygen supply. The genome sequence reveals gene clusters for three cytochrome c oxidases (ccoNOQP1, ccoNOQP2, and coxBA-coIII) and one quinol oxidase (cyoABCDE), all of which belong to the hemecopper superfamily (9,10,11). Heme-copper oxidases can be inhibited by cyanide, resulting in a block of electron transport via these oxidases.…”