2-heptyl-3-hydroxy-4-quinolone and homoserine-lactone

Bacteria use diverse small molecules for extra- and intracellular signaling. They scan small-molecule mixtures to access information about both their extracellular environment and their intracellular physiological status, and based on this information, they continuously interpret their circumstances and react rapidly to changes. Bacteria must integrate extra- and intracellular signaling information to mount appropriate responses to changes in their environment. We review recent research into two fundamental bacterial small-molecule signaling pathways: extracellular quorum-sensing signaling and intracellular cyclic dinucleotide signaling. We suggest how these two pathways may converge to control complex processes including multicellularity, biofilm formation, and virulence. We also outline new questions that have arisen from recent studies in these fields.

Topics: 4-Butyrolactone; Bacterial Physiological Phenomena; Bacterial Proteins; Biofilms; Cyclic GMP; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Genes, Bacterial; Homoserine; Lactones; Models, Biological; Oligopeptides; Phosphoric Diester Hydrolases; Phosphorus-Oxygen Lyases; Purine Nucleotides; Quinolones; Second Messenger Systems; Signal Transduction; Virulence

Many bacteria communicate with each other through signalling molecules, a process known as cell-to-cell communication. During this process, it is important for the signalling molecules to: (1) reach the target cells; and (2) to be received by the cognate receptor. Barriers such as the presence of extracellular matrix may prevent signals from reaching their targets; however, the influence of the extracellular matrix on cell-to-cell communication has scarcely been studied. Here, we demonstrate that the overproduction of an extracellular matrix, alginate, in a Pseudomonas aeruginosa mucoid variant, alters cell-to-cell communication by interfering with the response to quinolone signals while having no effect on N-acyl-L-homoserine lactones. The inhibition of quinolone signalling by alginate is limited to the alginate overproducer and has no effect on neighbour cells that do not produce alginate. Our study indicates that alginate overproduction affects the cell-to-cell communication of the mucoid variant, which may results in different downstream behaviours when it emerges in the presence of the wild-type (WT).

Topics: 4-Butyrolactone; Alginates; Bacterial Proteins; Extracellular Matrix; Gene Expression Regulation, Bacterial; Glucuronic Acid; Hexuronic Acids; Pseudomonas aeruginosa; Quinolones; Quorum Sensing; Signal Transduction

Quorum sensing inhibitors (QSIs) act as antivirulent agents since quorum sensing (QS) plays a vital role in regulating pathogenesis of Pseudomonas aeruginosa. However, application of single QSI may not be effective as pathogen is vulnerable to successful mutations. In such conditions, combination of QSIs can be exploited as there can be synergistic or adjuvant action. In the present study, we evaluated the antivirulence efficacy of combination of Vaccinium macrocarpon proanthocyanidin active fraction (PAF) and ciprofloxacin (CIP) at their sub-MICs using standard methods followed by analysis of their mode of action on QS using TLC and molecular docking. There was significant improvement in action of CIP when it was combined with PAF in reducing the QS controlled virulence factors (p < 0.05), motilities and biofilm of P. aeruginosa. TLC profiles of QS signals [(Acyl homoserine lactone (AHL) and Pseudomonas quinolone signal (PQS)] indicated that CIP in combination with PAF, besides showing inhibitory action on production of AHLs, also modulated production and inactivation of PQS. Docking scores also supported the observation. We therefore hypothesize that PAF-CIP combination, having improved anti-virulence property; can be exploited as a potent drug pairing against P. aeruginosa.

Topics: 4-Butyrolactone; Acyl-Butyrolactones; Adjuvants, Pharmaceutic; Anti-Bacterial Agents; Biofilms; Ciprofloxacin; Drug Synergism; Microbial Sensitivity Tests; Molecular Docking Simulation; Plant Extracts; Proanthocyanidins; Pseudomonas aeruginosa; Pseudomonas Infections; Quinolones; Quorum Sensing; Vaccinium macrocarpon; Virulence

In Pseudomonas aeruginosa the production of multiple virulence factors depends on cell-to-cell communication through the integration of N-acylhomoserine lactone (AHL)- and 2-heptyl-3-hydroxy-4(1H)-quinolone (PQS)- dependent signalling. Mutation of genes encoding the efflux protein MexI and the porin OpmD from the MexGHI-OpmD pump resulted in the inability to produce N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-c12-hsl) and pqs and a marked reduction in n-butanoyl-L-homoserine lactone levels. Both pump mutants were impaired in growth and exhibited enhanced rather than reduced antibiotic resistance. Provision of exogenous PQS improved growth and restored AHL and virulence factor production as well as antibiotic susceptibility, indicating that the pump mutants retained their capacity to respond to PQS. RT-PCR analysis indicated that expression of the PQS biosynthetic genes, phnA and pqsA, was inhibited when the mutants reached stationary phase, suggesting that the pleiotropic phenotype observed may be due to intracellular accumulation of a toxic PQS precursor. To explore this hypothesis, double mexI phnA (unable to produce anthranilate, the precursor of PQS) and mexI pqsA mutants were constructed; the improved growth of the former suggested that the toxic compound is likely to be anthranilate or a metabolite of it. Mutations in mexI and opmD also resulted in the attenuation of virulence in rat and plant infection models. In plants, addition of PQS restored the virulence of mexI and opmD mutants. Collectively, these results demonstrate an essential function for the MexGHI-OpmD pump in facilitating cell-to-cell communication, antibiotic susceptibility and promoting virulence and growth in P. aeruginosa.

Topics: 4-Butyrolactone; 4-Quinolones; Animals; Bacterial Outer Membrane Proteins; Base Sequence; DNA, Bacterial; Drug Resistance, Multiple, Bacterial; Female; Gene Expression; Genes, Bacterial; Lactuca; Membrane Transport Proteins; Mutation; Phenotype; Plant Diseases; Pseudomonas aeruginosa; Pseudomonas Infections; Quinolones; Rats; Rats, Inbred Lew; Signal Transduction; Virulence

Pseudomonas aeruginosa releases a spectrum of well-regulated virulence factors, controlled by intercellular communication (quorum sensing) and mediated through the production of small diffusible quorum-sensing signal molecules (QSSM). We hypothesize that QSSM may in fact serve a dual purpose, also allowing bacterial colonization via their intrinsic immune-modulatory capacity. One class of signal molecule, the N-acylhomoserine lactones, has pleiotropic effects on eukaryotic cells, particularly those involved in host immunity. In the present study, we have determined the comparative effects of two chemically distinct and endobronchially detectable QSSM, N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) and 2-heptyl-3-hydroxy-4 (1H)-quinolone or the Pseudomonas quinolone signal (PQS), on human leukocytes exposed to a series of stimuli designed to detect differential immunological activity in vitro. 3-Oxo-C12-HSL and PQS displayed differential effects on the release of interleukin-2 (IL-2) when human T cells were activated via the T-cell receptor and CD28 (a costimulatory molecule). 3-Oxo-C12-HSL inhibited cell proliferation and IL-2 release; PQS inhibited cell proliferation without affecting IL-2 release. Both molecules inhibited cell proliferation and the release of IL-2 following mitogen stimulation. Furthermore, in the presence of Escherichia coli lipopolysaccharide, 3-oxo-C12-HSL inhibited tumor necrosis factor alpha release from human monocytes, as reported previously (K. Tateda et al., Infect. Immun. 64:37-43, 1996), whereas PQS did not inhibit in this assay. These data highlight the presence of two differentially active immune modulatory QSSM from P. aeruginosa, which are detectable endobronchially and may be active at the host/pathogen interface during infection with P. aeruginosa, should the bronchial airway lymphoid tissues prove to be accessible to QSSM.

Topics: 4-Butyrolactone; Concanavalin A; Gene Expression Regulation; Homoserine; Humans; Interleukin-2; Leukocytes, Mononuclear; Lipopolysaccharides; Lymphocyte Activation; Pseudomonas aeruginosa; Quinolones; Signal Transduction; Tumor Necrosis Factor-alpha

Intrinsic and acquired antibiotic resistance of the nosocomial pathogen Pseudomonas aeruginosa is mediated mainly by the expression of several efflux pumps of broad substrate specificity. Here we report that nfxC type mutants, overexpressing the MexEF-OprN efflux system, produce lower levels of extracellular virulence factors than the susceptible wild type. These include pyocyanin, elastase, and rhamnolipids, three factors controlled by the las and rhl quorum-sensing systems of P. aeruginosa. In agreement with these observations are the decreased transcription of the elastase gene lasB and the rhamnosyltransferase genes rhlAB measured in nfxC type mutants. Expression of the lasR and rhlR regulator genes was not affected in the nfxC type mutant. In contrast, transcription of the C4-homoserine lactone (C4-HSL) autoinducer synthase gene rhlI was reduced by 50% in the nfxC type mutant relative to that in the wild type. This correlates with a similar decrease in C4-HSL levels detected in supernatants of the nfxC type mutant. Transcription of an rhlAB-lacZ fusion could be partially restored by the addition of synthetic C4-HSL and Pseudomonas quinolone signal (PQS). It is proposed that the MexEF-OprN efflux pump affects intracellular PQS levels.

Topics: 4-Butyrolactone; Anti-Bacterial Agents; Bacterial Outer Membrane Proteins; Bacterial Proteins; Base Sequence; Gene Expression Regulation, Bacterial; Hexosyltransferases; Ligases; Molecular Sequence Data; Mutation; Pancreatic Elastase; Pseudomonas aeruginosa; Quinolones; Sequence Analysis, DNA; Signal Transduction; Trans-Activators; Transcription Factors; Transcription, Genetic; Virulence