Quorum-sensing Mechanism Research Articles

Exposure to One Antibiotic Leads to Acquisition of Resistance to Another Antibiotic via Quorum Sensing Mechanisms.

The vancomycin-resistant Enterococci (VRE) have progressively become a severe medical problem. Although clinics have started to reduce vancomycin prescription, vancomycin resistance has not been contained. We found that the transfer of vancomycin resistance in Enterococcus faecalis increased more than 30-fold upon treatment by streptomycin. Notably, treatment with an antibiotic caused the bacteria to become resistant to another. The response was even stronger in the well-studied plasmid pCF10 and the number of transconjugants increased about 100,000-fold. We tested four different antibiotics, and all of them induced conjugal response. Through a mathematical model based on gene regulation, we found a plausible explanation. Via quorum sensing, the change of the cell density triggers the conjugation. Moreover, we searched for generality and found a similar strategy in Bacillus subtilis. The outcome of the present study suggests that even common antibiotics must not be overused.

Co-dependent and Interdigitated: Dual Quorum Sensing Systems Regulate Conjugative Transfer of the Ti Plasmid and the At Megaplasmid in Agrobacterium tumefaciens 15955.

Members of the Rhizobiaceae, often carry multiple secondary replicons in addition to the primary chromosome with compatible repABC-based replication systems. Unlike secondary chromosomes and chromids, repABC-based megaplasmids and plasmids can undergo copy number fluctuations and are capable of conjugative transfer in response to environmental signals. Several Agrobacterium tumefaciens lineages harbor three secondary repABC-based replicons, including a secondary chromosome (often linear), the Ti (tumor-inducing) plasmid and the At megaplasmid. The Ti plasmid is required for virulence and encodes a conjugative transfer (tra) system that is strictly regulated by a subset of plant-tumor released opines and a well-described acyl-homoserine lactone (AHL)-based quorum-sensing mechanism. The At plasmids are generally not required for virulence, but carry genes that enhance rhizosphere survival, and these plasmids are often conjugatively proficient. We report that the At megaplasmid of the octopine-type strain A. tumefaciens 15955 encodes a quorum-controlled conjugation system that directly interacts with the paralogous quorum sensing system on the co-resident Ti plasmid. Both the pAt15955 and pTi15955 plasmids carry homologs of a TraI-type AHL synthase, a TraR-type AHL-responsive transcription activator, and a TraM-type anti-activator. The traI genes from both pTi15955 and pAt15955 can direct production of the inducing AHL (3-octanoyl-L-homoserine lactone) and together contribute to the overall AHL pool. The TraR protein encoded on each plasmid activates AHL-responsive transcription of target tra gene promoters. The pAt15955 TraR can cross-activate tra genes on the Ti plasmid as strongly as its cognate tra genes, whereas the pTi15955 TraR is preferentially biased toward its own tra genes. Putative tra box elements are located upstream of target promoters, and comparing between plasmids, they are in similar locations and share an inverted repeat structure, but have distinct consensus sequences. The two AHL quorum sensing systems have a combinatorial effect on conjugative transfer of both plasmids. Overall, the interactions described here have implications for the horizontal transfer and evolutionary stability of both plasmids and, in a broad sense, are consistent with other repABC systems that often have multiple quorum-sensing controlled secondary replicons.

Molecular Mechanisms of Campylobacter Biofilm Formation and Quorum Sensing.

Even though Campylobacter spp. are known to be fastidious organisms, they can survive within the natural environment. One mechanism to withstand unfavourable conditions is the formation of biofilms, a multicellular structure composed of different bacterial and other microbial species which are embedded in an extracellular matrix. High oxygen levels, low substrate concentrations and the presence of external DNA stimulate the biofilm formation by C. jejuni. These external factors trigger internal adaptation processes, e.g. via regulating the expression of genes encoding proteins required for surface structure formation, as well as motility, stress response and antimicrobial resistance. Known genes impacting biofilm formation will be summarized in this review. The formation of biofilms as well as the expression of virulence genes is often regulated in a cell density depending manner by quorum sensing, which is mediated via small signalling molecules termed autoinducers. Even though quorum sensing mechanisms of other bacteria are well understood, knowledge on the role of these mechanisms in C. jejuni biofilm formation is still scarce. The LuxS enzyme involved in generation of autoinducer-2 is present in C. jejuni, but autoinducer receptors have not been identified so far. Phenotypes of C. jejuni strains lacking a functional luxS like reduced growth, motility, oxygen stress tolerance, biofilm formation, adhesion, invasion and colonization are also summarized within this chapter. However, these phenotypes are highly variable in distinct C. jejuni strains and depend on the culture conditions applied.

QUORUM SENSING in Mycobacterium tuberculosis. Review

SummaryThe rise in the incidence of tuberculosis expected by many phthisiatricians during the period of the covid pan-demic did not occur, its steady decline continues, which could be due to a sharp reduction in contacts. However, it cannot be ruled out that an extreme increase in stress-ful social factors associated withcovid pandemic may lead to an exacerbation of latent tuberculosis. In addi-tion, the growth of severe forms of the disease, caused by drug-resis tant strains of mycobacterium tuberculosis, continues. The drug resistance of the pathogen, which WHO has de signated as a global threat, the crisis of clas-sical antibio tic therapy dictate the need to develop a new strategy for the treatment of infectious diseases, which consists of influencing the genetic and enzymatic targets that determine the virulence of pathogens. One of the manifestations of bacterial adaptation as a component of virulence is QUORUM SENSING (QS) — a supra organic lev-el of self-organization of bacteria, due to which microbial communities can behave like a multicellular organism. The main phenomenon of QS is the formation of microbi-al biofilms — special communities of microorganisms en-closed in a biopolymer extracellular matrix, which serves as a direct obstacle to the action of immunocompetent cells and antibacterial substances, and also increases drug resistance many times over. Myco bacterial biofilm shave been discovered, which play an essential role in the formation of case ous necrosis, decay cavities and drug resistance. In recent years, there has been an active search for QS inhibitors as tools for alternative therapeu-tic interventions in infectious pathology. In publications on mycobacterial films, it is also emphasized that the dis-covered QS mechanisms in MBT are promising therapeu-tic targets. Thus, the problem of QS of mycobacterium tu-berculosis requires further in-depth and comprehensive study in order to develop methods of alternative therapy for tuberculosis.

Function of quorum sensing and cell signaling in wastewater treatment systems.

Quorum sensing (QS) is a communication mode between microorganisms to regulate bacteria ecological relations and physiological behaviors, thus achieve the physiological function that single bacteria cannot complete. This phenomenon plays important roles in the formation of biofilm and granular sludge, and may be related to enhancement of some functional bacteria activity in wastewater treatment systems. There is a need to better understand bacterial QS in engineered reactors, and to assess how designs and operations might improve the removal efficiency. This article reviewed the recent advances of QS in several environmental systems and mainly analyzed the regulation mechanism of QS-based strategies for biofilm, granular sludge, functional bacteria, and biofouling control. The co-existences of multiple signal molecules in wastewater treatment (WWT) processes were also summarized, which provide basis for the future research on the QS mechanism of multiple signal molecules' interaction in WWT. This review would present some prospects and suggestions which are of practical significance for further application.

Anti-quorum Sensing and Protective Efficacies of Naringin Against Aeromonas hydrophila Infection in Danio rerio.

It is now well known that the quorum sensing (QS) mechanism coordinates the production of several virulence factors and biofilm formation in most pathogenic microorganisms. Aeromonas hydrophila is a prime pathogen responsible for frequent outbreaks in aquaculture settings. Recent studies have also continuously reported that A. hydrophila regulates virulence factor production and biofilm formation through the QS system. In addition to the presence of antibiotic resistance genes, biofilm-mediated antibiotic resistance increases the severity of A. hydrophila infections. To control the bacterial pathogenesis and subsequent infections, targeting the QS mechanism has become one of the best alternative methods. Though very few compounds were identified as QS inhibitors against A. hydrophila, to date, the screening and identification of new and effective natural QS inhibitors is a dire necessity to control the infectious A. hydrophila. The present study endorses naringin (NA) as an anti-QS and anti-infective agent against A. hydrophila. Initially, the NA showed a concentration-dependent biofilm reduction against A. hydrophila. Furthermore, the results of microscopic analyses and quantitative virulence assays displayed the promise of NA as a potential anti-QS agent. Subsequently, the downregulation of ahh1, aerA, lip and ahyB validate the interference of NA in virulence gene expression. Furthermore, the in vivo assays were carried out in zebrafish model system to evaluate the anti-infective potential of NA. The outcome of the immersion challenge assay showed that the recovery rate of the zebrafish has substantially increased upon treatment with NA. Furthermore, the quantification of the bacterial load upon NA treatment showed a decreased level of bacterial counts in zebrafish when compared to the untreated control. Moreover, the NA treatment averts the pathogen-induced histoarchitecture damages in vital organs of zebrafish, compared to their respective controls. The current study has thus analyzed the anti-QS and anti-infective capabilities of NA and could be employed to formulate effective treatment measures against A. hydrophila infections.

The Roles of Microbial Cell-Cell Chemical Communication Systems in the Modulation of Antimicrobial Resistance.

Rapid emergence of antimicrobial resistance (AMR) has become a critical challenge worldwide. It is of great importance to understand how AMR is modulated genetically in order to explore new antimicrobial strategies. Recent studies have unveiled that microbial communication systems, which are known to play key roles in regulation of bacterial virulence, are also associated with the formation and regulation of AMR. These microbial cell-to-cell chemical communication systems, including quorum sensing (QS) and pathogen–host communication mechanisms, rely on detection and response of various chemical signal molecules, which are generated either by the microbe itself or host cells, to activate the expression of virulence and AMR genes. This article summarizes the generic signaling mechanisms of representative QS and pathogen–host communications systems, reviews the current knowledge regarding the roles of these chemical communication systems in regulation of AMR, and describes the strategies developed over the years for blocking bacterial chemical communication systems in disease control. The research progress in this field suggests that the bacterial cell-cell communication systems are a promising target not only for disease control but also for curbing the problem of microbial drug resistance.

Functional characterization of quorum sensing LuxR-type transcriptional regulator, EasR in Enterobacter asburiae strain L1.

Over the past decades, Enterobacter spp. have been identified as challenging and important pathogens. The emergence of multidrug-resistant Enterobacteria especially those that produce Klebsiella pneumoniae carbapenemase has been a very worrying health crisis. Although efforts have been made to unravel the complex mechanisms that contribute to the pathogenicity of different Enterobacter spp., there is very little information associated with AHL-type QS mechanism in Enterobacter spp. Signaling via N-acyl homoserine lactone (AHL) is the most common quorum sensing (QS) mechanism utilized by Proteobacteria. A typical AHL-based QS system involves two key players: a luxI gene homolog to synthesize AHLs and a luxR gene homolog, an AHL-dependent transcriptional regulator. These signaling molecules enable inter-species and intra-species interaction in response to external stimuli according to population density. In our recent study, we reported the genome of AHL-producing bacterium, Enterobacter asburiae strain L1. Whole genome sequencing and in silico analysis revealed the presence of a pair of luxI/R genes responsible for AHL-type QS, designated as easI/R, in strain L1. In a QS system, a LuxR transcriptional protein detects and responds to the concentration of a specific AHL controlling gene expression. In E. asburiae strain L1, EasR protein binds to its cognate AHLs, N-butanoyl homoserine lactone (C4-HSL) and N–hexanoyl homoserine lactone (C6-HSL), modulating the expression of targeted genes. In this current work, we have cloned the 693 bp luxR homolog of strain L1 for further characterization. The functionality and specificity of EasR protein in response to different AHL signaling molecules to activate gene transcription were tested and validated with β-galactosidase assays. Higher β-galactosidase activities were detected for cells harboring EasR, indicating EasR is a functional transcriptional regulator. This is the first report documenting the cloning and characterization of transcriptional regulator, luxR homolog of E. asburiae.

Recent Breakthroughs in Various Antimicrobial Resistance Induced Quorum Sensing Biosynthetic Pathway Mediated Targets and Design of their Inhibitors

The world is under the grasp of dangerous post-antibiotics and antimicrobials attack where common infections may become untreatable, leading to premature deaths due to antimicrobial resistance (AMR). While an estimated 7,00,000 people die annually due to AMR, which is a public health threat to all communities in different parts of the world regardless of their economic status; however, this threat is serious in low- and middle-income countries having lack of sanitation and health infrastructure. The 68th World Health Assembly endorsed the Global Action Plan on antimicrobial resistance. Consequently, many countries started drafting and committing to National Action Plans against AMR. As strong as National Action Plans are in terms of prescribing rational use of antimicrobials, infection control practices, and related public health measures, without strong healthcare systems, these measures will have a limited impact on AMR in developing countries. The major reason for AMR is microbial quorum sensing (QS) that may strengthen the microbial community to generate inter-communication and virulence effects via quorum sensing mechanisms. Global stewardship to combat antimicrobial resistance aims to develop anti-quorum sensing compounds that can inhibit the biosynthetic pathway mediated different quorum sensing targets. It may pave an effective attempt to minimize microbial quorum sensing mediated antimicrobial resistance. The present review describes QS mediated various potential target enzymes, their connection to AMR, and finds out the corresponding QS biosynthetic target inhibitors. These potential inhibitors can be derivatized to design and develop next-generation antimicrobial agents.

Terminalia chebula and Ficus racemosa principles mediated repression of novel drug target Las R – the transcriptional regulator and its controlled virulence factors produced by multiple drug resistant Pseudomonas aeruginosa - Biocompatible formulation against drug resistant bacteria

Pseudomonas aeruginosa– major group of an aerobic bacteria associated with nosocomial and other life threatening infections. Diverse virulence factors produced by P. aeruginosa is due to distinct molecular cell signaling mechanism termed as quorum sensing (QS). Interfering with normal QS mechanism by active biomolecules is an effective strategy for attenuating its virulence. With this objective, the present study is undertaken to evaluate the inhibition of quorum sensing of clinical isolate of P. aeruginosa by repression of Las R-a transcriptional regulator for QS by ethanol extract of Terminalia chebula and Ficus racemosa. Las R repression by the plant extracts was measured in inhibition of various virulence factors like biofilm, pyocyanin production, total proteolytic activity, swarming and twisting motility. Fabrication of the extracted metabolites on the wound dressing and its effect on anti bacterial activity was also investigated. Compatibility of plant extracts on zebra fish development and blood cells was further studied. P. aeruginosa was isolated from the post operative patient and the isolated pure culture was identified by cultural, biochemical, molecular characteristics. Active principles of both the plants were readily extracted in ethanol and effectively repressed the expression of Las R. Both the tested plant extracts effectively repressed Las R expression which in turn affect the production of various virulence factors like biofilm formation, pyocyanin production, swarming motility, twisting motility, total proteolytic activity, cell adhesion and signaling molecule acyl honoserine lactone (AHL) production. Plant extract treatment brought about drastic reduction of all the tested virulence factors and AHL production. Extracted metabolites were fabricated on the wound dressing material adopting simple dip or immersion method reveals uniform coating, effective embedding of phytochemicals with the fibers and retained the anti bacterial activity against P. aeruginosa. Biocompatibility studies with zebra fish model shows both the tested plant extracts treatment was not exhibited any sign of toxicity on the developmental stages of Zebra fish. Hemolysis and changes in anti oxidative enzymes were not recorded in the plant extracts treated blood which demonstrated the best biocompatibility of the tested plant extracts. These results shows that the presence of potential phytochemicals in the ethanolic extract of Terminalia chebula and Ficus racemosa effectively represses the Las R followed by inhibition of quorum sensing mediated virulence factors production may be useful in the lead of anti bacterial drugs.

Genome-Scale Metabolic Model of Xanthomonas phaseoli pv. manihotis: An Approach to Elucidate Pathogenicity at the Metabolic Level.

Xanthomonas phaseoli pv. manihotis (Xpm) is the causal agent of cassava bacterial blight, the most important bacterial disease in this crop. There is a paucity of knowledge about the metabolism of Xanthomonas and its relevance in the pathogenic process, with the exception of the elucidation of the xanthan biosynthesis route. Here we report the reconstruction of the genome-scale model of Xpm metabolism and the insights it provides into plant–pathogen interactions. The model, iXpm1556, displayed 1,556 reactions, 1,527 compounds, and 890 genes. Metabolic maps of central amino acid and carbohydrate metabolism, as well as xanthan biosynthesis of Xpm, were reconstructed using Escher (https://escher.github.io/) to guide the curation process and for further analyses. The model was constrained using the RNA-seq data of a mutant of Xpm for quorum sensing (QS), and these data were used to construct context-specific models (CSMs) of the metabolism of the two strains (wild type and QS mutant). The CSMs and flux balance analysis were used to get insights into pathogenicity, xanthan biosynthesis, and QS mechanisms. Between the CSMs, 653 reactions were shared; unique reactions belong to purine, pyrimidine, and amino acid metabolism. Alternative objective functions were used to demonstrate a trade-off between xanthan biosynthesis and growth and the re-allocation of resources in the process of biosynthesis. Important features altered by QS included carbohydrate metabolism, NAD(P)+ balance, and fatty acid elongation. In this work, we modeled the xanthan biosynthesis and the QS process and their impact on the metabolism of the bacterium. This model will be useful for researchers studying host–pathogen interactions and will provide insights into the mechanisms of infection used by this and other Xanthomonas species.

ACYL HOMOSERINE LACTONE LACTONASE BACTERIA POTENTIAL AS BIOCONTROL AGENT OF SOFT ROT INFECTION

The virulence gene expression of phytopathogenic bacteria, such as Dickeya dadantii is regulated by quorum sensing (QS) mechanism. The QS mechanism is induced by acyl homoserine lactone (AHL) that is universal autoinducer of Gram-negative bacteria. Degradation of AHL compound can be used to control the virulence of phytopathogenic bacteria. The enzyme of AHL lactonase is encoded by the aiiA gene and can degrade the lactone ring of AHL. This study was aimed to characterize aiiA gene encoding AHL lactonase from 20 isolates of shrimp pond and soil forest, Indonesia and examine the isolates ability to reduce violacein pigment of Chromobacterium violaceum and soft rot disease symptoms caused by D. dadantii. The results showed nine bacterial isolates (B4, B10, B11, B13, B14, B16, B17, B18 and B19) containing the aiiA gene encoding AHL lactonase. Based on 16S rRNA gene identification, 9 isolates had the highest similarity with Bacillus toyonensis, B.cereus, B. thuringiensis, B. mycoides and B. subtilis. These isolates had the capability to reduce the formation of violacein pigment and the symptoms of soft rot disease in potatoes. These results show that the AHL lactonase produced by these isolates can be used as Quorum Quenching (QQ) agent to inhibit the QS mechanism

Acyl-Homoserine Lactone from Plant-Associated Pseudomonas sp. Influences Solanum lycopersicum Germination and Root Growth.

It is known that plant and associated bacteria coevolved, but just now the roles of chemical signaling compounds in these intricate relationships have been systematically studied. Many Gram-negative bacteria produce N-acyl-L-homoserine lactones (AHL), chemical signals used in quorum-sensing bacterial communications mechanisms. In recent years, it has been shown that these compounds may also influence the development of plants, acting as allelochemicals, in still not well understood eukaryot-prokaryot interactions. In the present work, a quorum-sensing molecule produced by the tomato associated bacterium Pseudomonas sp. was characterized and its effects on germination and growth of tomato seedlings were accessed. The chemical study of the bacterium extract led to the identification of N-3-oxo-dodecanoyl-L-homoserine lactone (1), using gas chromatography coupled to electron impact mass spectrometry (GC-MS), and ultra-high resolution Qq-time-of-flight mass spectrometry (UHR-QqTOF-MS) equipped with an electrospray ionization source (ESI). The synthetic compound was tested at different concentrations in tomato to evaluate its effects on seed germination and seedlings root growth. Inhibition of tomato seed germination and root growth were observed in the presence of micromolar concentrations of the compound 1. Scanning electron microscopy evidenced morphological alterations on roots in the presence of thecompound, with reduction of growth, impaired root hairs development and cracks in the rhizodermis.

Farnesol and Tyrosol: Secondary Metabolites with a Crucial quorum-sensing Role in Candida Biofilm Development.

When living in biological and interactive communities, microorganisms use quorum-sensing mechanisms for their communication. According to cell density, bacteria and fungi can produce signaling molecules (e.g., secondary metabolites), which participate, for example, in the regulation of gene expression and coordination of collective behavior in their natural niche. The existence of these secondary metabolites plays a main role in competence, colonization of host tissues and surfaces, morphogenesis, and biofilm development. Therefore, for the design of new antibacterials or antifungals and understanding on how these mechanisms occur, to inhibit the secretion of quorum-sensing (e.g., farnesol and tyrosol) molecules leading the progress of microbial infections seems to be an interesting option. In yeasts, farnesol has a main role in the morphological transition, inhibiting hyphae production in a concentration-dependent manner, while tyrosol has a contrary function, stimulating transition from spherical cells to germ tube form. It is beyond doubt that secretion of both molecules by fungi has not been fully described, but specific meaning for their existence has been found. This brief review summarizes the important function of these two compounds as signaling chemicals participating mainly in Candida morphogenesis and regulatory mechanisms.

Quorum quenching acylase impacts the viability and morphological change of Agrobacterium tumefaciens cells

Acylase is known as a quorum quenching enzyme that degrades N-acyl-homoserine lactones (AHLs), a key signaling molecule in a quorum sensing (QS) mechanism. Acylase I cleaves the acyl-chain in the chemical structures of AHLs, thereby exerting an anti-biofilm effect by the inhibition of bacterial cell-cell communication and resultant secretion of extracellular polymeric substances (EPS). However, the physical and physiological impacts of acylase on bacterial cells remain to be systematically elucidated. This study, therefore, investigated the effect of active and inactive acylase addition on the growth, viability, and cell morphologies of Agrobacterium tumefaciens. For comparison, active and inactive lysozymes were taken as positive controls. The results showed that active acylase inhibited A.tumefaciens cell growth at concentrations ranging from 0.1 to 1000μgmL-1, and so did active lysozyme. Fluorescent detection by Live/Dead staining underpinned that cell viability of A.tumefaciens decreased at concentrations higher than 0.1μgmL-1 for both acylase and lysozyme, although lysozyme inflicted higher degree of cellular damage. Moreover, atomic force microscopy unraveled a noticeable distortion of A.tumefaciens cells by both acylase and lysozyme. Together, the results showed that acylase not only blocked AHLs-based QS mechanisms but also compromised cell viability and altered surface morphology of A.tumefaciens cells, as observed by the addition of hydrolase.

Effect of Quorum Sensing Inhibitor Agents against Pseudomonas aeruginosa

The abuse of antibiotics in therapy has lead to the development of resistance in the target organisms. The failure of presented antibiotics to control infections makes it essential to discover option to presently available drugs. Quorum sensing (QS) is a used by many bacteria to regulate gene expression in accordance with population density through the use of signal molecules or autoinducers. The QS is used by Bacteria populations to communicate and coordinate their group interactions, which is applied by pathogens in infection processes. The QS pathways in bacteria are composed of several parts, including bacteria populations, signal molecules, protein activators and target genes. The pathogenicity in numerous bacteria is regulated by QS signaling systems. The QS inhibition system may cause the reduction of virulence and defense against bacterial infections. The QS is the main regulator of virulence and biofilm formation in Pseudomonas aeruginosa and other relevant bacteria. In P. aeruginosa, the expression of many virulence factors appears to be controlled by QS. So, according to the role of this mechanism in the regulation and production of many virulence factors, the function of QS is required for P. aeruginosa to cause disease and infection. In this article, we discussed the QS mechanism in gram-negative and gram-positive bacteria with a closer look at the P. aeruginosa. A variety of plants showed their effects on P. aeruginosa virulence. Extract of various plants control the regulatory QS genes and factors with marginal effects on bacterial growth. The quorum-quenching (QQ) mechanisms are unrelated to static or cidal effects. In fact, anti-QS have already shown promise in the battle against P. aeruginosa infections.

The extreme plant‐growth‐promoting properties ofPantoea phytobeneficialisMSR2 revealed by functional and genomic analysis

Numerous Pantoea strains are important because of the benefit they provide in the facilitation of plant growth. However, Pantoea have a high level of genotypic diversity and not much is understood regarding their ability to function in a plant beneficial manner. In the work reported here, the plant growth promotion activities and the genomic properties of the unusual Pantoea phytobeneficialis MSR2 are elaborated, emphasizing the genetic mechanisms involved in plant colonization and growth promotion. Detailed analysis revealed that strain MSR2 belongs to a rare group of Pantoea strains possessing an astonishing number of plant growth promotion genes, including those involved in nitrogen fixation, phosphate solubilization, 1-aminocyclopropane-1-carboxylic acid deaminase activity, indoleacetic acid and cytokinin biosynthesis, and jasmonic acid metabolism. Moreover, the genome of this bacterium also contains genes involved in the metabolism of lignin and other plant cell wall compounds, quorum-sensing mechanisms, metabolism of plant root exudates, bacterial attachment to plant surfaces and resistance to plant defences. Importantly, the analysis revealed that most of these genes are present on accessory plasmids that are found within a small subset of Pantoea genomes, reinforcing the idea that Pantoea evolution is largely mediated by plasmids, providing new insights into the evolution of beneficial plant-associated Pantoea.

Design, Synthesis, and Biochemical Characterization of Non-Native Antagonists of the Pseudomonas aeruginosa Quorum Sensing Receptor LasR with Nanomolar IC50 Values.

Quorum sensing (QS), a bacterial cell-to-cell communication system mediated by small molecules and peptides, has received significant interest as a potential target to block infection. The common pathogen Pseudomonas aeruginosa uses QS to regulate many of its virulence phenotypes at high cell densities, and the LasR QS receptor plays a critical role in this process. Small molecule tools that inhibit LasR activity would serve to illuminate its role in P. aeruginosa virulence, but we currently lack highly potent and selective LasR antagonists, despite considerable research in this area. V-06-018, an abiotic small molecule discovered in a high-throughput screen, represents one of the most potent known LasR antagonists but has seen little study since its initial report. Herein, we report a systematic study of the structure-activity relationships (SARs) that govern LasR antagonism by V-06-018. We synthesized a focused library of V-06-018 derivatives and evaluated the library for bioactivity using a variety of cell-based LasR reporter systems. The SAR trends revealed by these experiments allowed us to design probes with 10-fold greater potency than that of V-06-018 and 100-fold greater potency than other commonly used N-acyl-l-homoserine lactone (AHL)-based LasR antagonists, along with high selectivities for LasR. Biochemical experiments to probe the mechanism of antagonism by V-06-018 and its analogues support these compounds interacting with the native ligand-binding site in LasR and, at least in part, stabilizing an inactive form of the protein. The compounds described herein are the most potent and efficacious antagonists of LasR known and represent robust probes both for characterizing the mechanisms of LuxR-type QS and for chemical biology research in general in the growing QS field.

Revealing the proteomic profiles of antianthracnose-related proteins in Serratia plymuthica UBCR_12

Capacity of Serratia plymuthica UBCR_12 in suppressing Colletotrichum gloeosporioides is regulated molecularly through protein expression. Efficacy of this bacteria is greatly affected by various environmental factors. This study was aimed to investigate the expression profile of antianthracnose-related proteins in S. plymuthica UBCR_12 under different modified medium. Antagonistic activity against C. gloeosporioides was assessed using agar spot method in PDA medium supplemented with several concentrations of peptone or glucose. Co-culture was performed under the same condition as an antagonistic assay to induce the extracellular proteomes expression related to this antianthracnose trait. Proteome profiles were visualized using SDS-PAGE and 2-DE, then a highly differential protein spot was further analyzed using MALDI-TOF-MS. Prediction of protein classification and possible hypothetical pathway were performed in-silico. The highest suppression (42.3% at 9th-day post application) resulted in 2% glucose-supplemented medium, however 2% peptone-supplemented medium conferred a quicker suppression (at 5th-day post application) with quite lower inhibition efficacy (40%). Differential expression of 42 kDa protein band recorded during peptone addition was predicted to be flagellin protein, which might correlate with a rapid stimulus of suppression activity. This protein involved in the quorum-sensing mechanism by triggering the greater rate of cell division resulting in bacterial colonization and motility approaching the fungal pathogen.

Bacterial Second Messenger Cyclic di-AMP Modulates the Competence State in Streptococcus pneumoniae.

Streptococcus pneumoniae (the pneumococcus) is a naturally competent organism that causes diseases such as pneumonia, otitis media, and bacteremia. The essential bacterial second messenger cyclic di-AMP (c-di-AMP) is an emerging player in the stress responses of many pathogens. In S. pneumoniae, c-di-AMP is produced by a diadenylate cyclase, CdaA, and cleaved by phosphodiesterases Pde1 and Pde2. c-di-AMP binds a transporter of K+ (Trk) family protein, CabP, which subsequently halts K+ uptake via the transporter TrkH. Recently, it was reported that Pde1 and Pde2 are essential for pneumococcal virulence in mouse models of disease. To elucidate c-di-AMP-mediated transcription that may lead to changes in pathogenesis, we compared the transcriptomes of wild-type (WT) and Δpde1 Δpde2 strains by transcriptome sequencing (RNA-Seq) analysis. Notably, we found that many competence-associated genes are significantly upregulated in the Δpde1 Δpde2 strain compared to the WT. These genes play a role in DNA uptake, recombination, and autolysis. Competence is induced by a quorum-sensing mechanism initiated by the secreted factor competence-stimulating peptide (CSP). Surprisingly, the Δpde1 Δpde2 strain exhibited reduced transformation efficiency compared to WT bacteria, which was c-di-AMP dependent. Transformation efficiency was also directly related to the [K+] in the medium, suggesting a link between c-di-AMP function and the pneumococcal competence state. We found that a strain that possesses a V76G variation in CdaA produced less c-di-AMP and was highly susceptible to CSP. Deletion of cabP and trkH restored the growth of these bacteria in medium with CSP. Overall, our study demonstrates a novel role for c-di-AMP in the competence program of S. pneumoniaeIMPORTANCE Genetic competence in bacteria leads to horizontal gene transfer, which can ultimately affect antibiotic resistance, adaptation to stress conditions, and virulence. While the mechanisms of pneumococcal competence signaling cascades have been well characterized, the molecular mechanism behind competence regulation is not fully understood. The bacterial second messenger c-di-AMP has previously been shown to play a role in bacterial physiology and pathogenesis. In this study, we provide compelling evidence for the interplay between c-di-AMP and the pneumococcal competence state. These findings not only attribute a new biological function to this dinucleotide as a regulator of competence, transformation, and survival under stress conditions in pneumococci but also provide new insights into how pneumococcal competence is modulated.