Messing with Bacterial Quorum Sensing

Quorum sensing is widely recognized as an efficient mechanism to regulate expression of specific genes responsible for communal behavior in bacteria. Several bacterial phenotypes essential for the successful establishment of symbiotic, pathogenic, or commensal relationships with eukaryotic hosts, including motility, exopolysaccharide production, biofilm formation, and toxin production, are often regulated by quorum sensing. Interestingly, eukaryotes produce quorum-sensing-interfering (QSI) compounds that have a positive or negative influence on the bacterial signaling network. This eukaryotic interference could result in further fine-tuning of bacterial quorum sensing. Furthermore, recent work involving the synthesis of structural homologs to the various quorum-sensing signal molecules has resulted in the development of additional QSI compounds that could be used to control pathogenic bacteria. The creation of transgenic plants that express bacterial quorum-sensing genes is yet another strategy to interfere with bacterial behavior. Further investigation on the manipulation of quorum-sensing systems could provide us with powerful tools against harmful bacteria.     

L-Canavanine made by Medicago sativa interferes with quorum sensing in Sinorhizobium meliloti

Sinorhizobium meliloti is a gram-negative soil bacterium, capable of establishing a nitrogen-fixing symbiosis with its legume host, alfalfa (Medicago sativa). Quorum sensing plays a crucial role in this symbiosis, where it influences the nodulation process and the synthesis of the symbiotically important exopolysaccharide II (EPS II). S. meliloti has three quorum-sensing systems (Sin, Tra, and Mel) that use N-acyl homoserine lactones as their quorum-sensing signal molecule. Increasing evidence indicates that certain eukaryotic hosts involved in symbiotic or pathogenic relationships with gram-negative bacteria produce quorum-sensing-interfering (QSI) compounds that can cross-communicate with the bacterial quorum-sensing system. Our studies of alfalfa seed exudates suggested the presence of multiple signal molecules capable of interfering with quorum-sensing-regulated gene expression in different bacterial strains. In this work, we choose one of these QSI molecules (SWI) for further characterization. SWI inhibited violacein production, a phenotype that is regulated by quorum sensing in Chromobacterium violaceum. In addition, this signal molecule also inhibits the expression of the S. meliloti exp genes, responsible for the production of EPS II, a quorum-sensing-regulated phenotype. We identified this molecule as l-canavanine, an arginine analog, produced in large quantities by alfalfa and other legumes.     

Exploiting Quorum Sensing To Confuse Bacterial Pathogens

SUMMARY

Cell-cell communication, or quorum sensing, is a widespread phenomenon in bacteria that is used to coordinate gene expression among local populations. Its use by bacterial pathogens to regulate genes that promote invasion, defense, and spread has been particularly well documented. With the ongoing emergence of antibiotic-resistant pathogens, there is a current need for development of alternative therapeutic strategies. An antivirulence approach by which quorum sensing is impeded has caught on as a viable means to manipulate bacterial processes, especially pathogenic traits that are harmful to human and animal health and agricultural productivity. The identification and development of chemical compounds and enzymes that facilitate quorum-sensing inhibition (QSI) by targeting signaling molecules, signal biogenesis, or signal detection are reviewed here. Overall, the evidence suggests that QSI therapy may be efficacious against some, but not necessarily all, bacterial pathogens, and several failures and ongoing concerns that may steer future studies in productive directions are discussed. Nevertheless, various QSI successes have rightfully perpetuated excitement surrounding new potential therapies, and this review highlights promising QSI leads in disrupting pathogenesis in both plants and animals.     

Different aspects of bacterial communication signals

The communication or quorum-sensing signal molecules (QSSM) are specialized molecules used by numerous gram-negative bacterial pathogens of animals and plants to regulate or modulate bacterial virulence factor production. In plant-associated bacteria, genes encoding the production of these signal molecules, QSSMs, were discovered to be linked with the phenotype of bacterium, because mutation of these genes typically disrupts some behaviors of bacteria. There are other regulator genes which respond to the presence of signal molecule and regulate the production of signal molecule as well as some virulence factors. The synthesis and regulator genes (collectively called quorum-sensing genes hereafter) are repressed in low bacterial population but induced when bacteria reach to high cell density. Multiple regulatory components have been identified in the bacteria that are under control of quorum sensing. This review describes different communication signal molecules, and the various chemical, physical and genomic factors known to synthesize signals. Likewise, the role of some signal-degrading enzymes or compounds and the interaction of QSSMs with eukaryotic metabolism will be discussed here.     

The role of a periplasmic gluconolactonase (PpgL)-like protein in <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">syringae</Emphasis> B728a

The communication or quorum-sensing signal molecules (QSSM) are specialized molecules used by numerous gram-negative bacterial pathogens of animals and plants to regulate or modulate bacterial virulence factor production. In plant-associated bacteria, genes encoding the production of these signal molecules, QSSMs, were discovered to be linked with the phenotype of bacterium, because mutation of these genes typically disrupts some behaviors of bacteria. There are other regulator genes which respond to the presence of signal molecule and regulate the production of signal molecule as well as some virulence factors. The synthesis and regulator genes (collectively called quorum-sensing genes hereafter) are repressed in low bacterial population but induced when bacteria reach to high cell density. Multiple regulatory components have been identified in the bacteria that are under control of quorum sensing. This review describes different communication signal molecules, and the various chemical, physical and genomic factors known to synthesize signals. Likewise, the role of some signal-degrading enzymes or compounds and the interaction of QSSMs with eukaryotic metabolism will be discussed here.     

群体感应抑制剂对海洋生态功能菌生物膜形成的影响

[目的]研究天然群体感应抑制剂(Quorum sensing inhibitors,QSI)分子对海洋生态功能菌生物膜形成的影响.[方法]以对污损生物幼虫附着具有诱导作用的海洋细菌为目标菌,通过在其生物膜的形成过程中添加天然群体感应抑制剂,研究其对目标菌成膜细菌数和浮游细菌数、生物膜形态以及生物膜表面胞外多糖含量的影响.[结果]呋喃酮和吡啶在50 mg/L时,对8株目标菌的成膜有显著的抑制作用,抑制率在80％左右,吲哚、青霉烷酸和香豆素在较高浓度800 mg/L才有比较好的抑制活性.生长抑制实验结果显示,同等浓度下,QSI分子对目标菌成膜的抑制活性明显高于其对浮游细菌生长的抑制活性.结果表明,QSI分子主要通过干扰目标菌群体感应系统以抑制生物膜的形成.[结论]研究证实QSI分子在海洋菌生物膜形成过程中具有一定的调控作用.通过添加QSI可能能够间接抑制由生物膜诱导的污损生物附着,从而以新的角度研制新型抗污损物质.     

植物精油对铜绿假单胞菌群体感应系统的抑制作用研究进展

群体感应（quorum sensing,QS）是细菌个体与个体之间的一种交流机制,广泛存在于细菌中。铜绿假单胞菌是人类的一种条件致病菌,它具有至少3种QS系统,即las、rhl和pqs系统,且各系统之间存在着级联调控关系,它们共同作用调控着该菌众多毒力基因的表达和毒力因子的产生。近年来,通过抑制铜绿假单胞菌的QS系统以控制其毒力和致病力,成为一种新型的铜绿假单胞菌感染防控策略。植物精油是一种天然的群体感应抑制剂（quorum sensing inhibitors, QSI）,多种精油活性化合物都能抑制铜绿假单胞菌的QS系统,而且尚未发现细菌对其产生耐药性。基于此,梳理了铜绿假单胞菌QS系统的组成及其级联调控关系,简要介绍了植物精油的QS抑制机制和抑制活性,并重点综述了萜烯类化合物、芳香族化合物、脂肪族化合物、含硫含氮化合物4类精油化合物对铜绿假单胞菌QS系统抑制作用的研究进展,以期为从天然化合物中发现和筛选安全、高效的细菌QSI的相关研究提供参考,并为致病菌的防控奠定理论基础。     

真菌中的群体感应系统

以胞间通讯信号分子介导的细菌群体感应参与细菌多种生理功能的调控是非常普遍的。近年的研究表明,真菌中也存在类似于细菌群体感应信号分子的调节分子,并且介导着真菌某些生理行为的调节。这一过程也称为真菌的群体感应系统。文中简要介绍真菌群体感应系统的研究进展,并讨论了真菌群体感应系统作为抗真菌感染靶点的可能性。     

群体感应抑制剂的筛选及其在微生物病害防治中的应用

在绝大多数病原菌中都发现有群体感应机制存在,用于调控侵染过程中微生物致病基因的表达。群体感应抑制剂处理既能控制微生物致病毒性,又不影响细胞生长,因此不会导致抗性株的形成,是一种理想的抗病原性药物。本文重点探讨了群体感应抑制剂的筛选、种类以及在群体感应过程中的作用机理和潜在应用价值。     

Bacterial social engagements

Quorum sensing is a process that enables bacteria to communicate using secreted signaling molecules called autoinducers. This process enables a population of bacteria to regulate gene expression collectively and, therefore, control behavior on a community-wide scale. Quorum sensing is widespread in the bacterial world and, generally, processes controlled by quorum sensing are unproductive when undertaken by an individual bacterium but become effective when undertaken by the group. Cell-cell communication can occur within and between bacterial species, and between bacteria and their eukaryotic hosts, which suggests that the chemical lexicon is complex. Prokaryotic and eukaryotic mechanisms for enhancing and inhibiting quorum sensing have been identified, which suggests that manipulation of quorum-sensing-controlled processes could be common in bacterial-bacterial and bacterial-eukaryotic associations.     

Presence of quorum-sensing inhibitor-like compounds from bacteria isolated from the brown alga Colpomenia sinuosa

Aims:  Several Gram-negative bacterial species use N -acyl homoserine lactone (AHL) molecules as quorum-sensing (QS) signals to regulate various biological functions. Similarly, various bacteria can stimulate, inhibit or inactivate QS signals in other bacteria by producing molecules called as quorum-sensing inhibitors (QSI). Our aim was to screen and identify the epibiotic bacteria associated with brown algae for their ability of producing QS-inhibiting activity.
Methods and Results:  QSI screenings were conducted on several epibiotic bacteria isolated from a marine brown alga Colpomenia sinuosa , using Serratia rubidaea JCM 14263 as an indicator organism. Strain JCM 14263 controls the production of red pigment, prodigiosin by AHL QS. Out of 96 bacteria, which were isolated from the surface of the brown alga, 12% of strains showed the ability to produce QSI, which was observed from the pigmentation inhibition on Ser. rubidaea JCM 14263 without affecting its growth. Phylogenetic analysis using 16S rRNA gene sequencing method demonstrated bacterial isolates showing QS inhibition-producing bacteria belonging to the Bacillaceae (Firmicutes), Pseudomonadaceae (Proteobacteria), Pseudoalteromonadaceae (Proteobacteria) and Vibrionaceae (Proteobacteria).
Conclusion:  An appreciable percentage of bacteria isolated from the brown alga produced QSI-like compounds.
Significance and Impact of the Study:  The screening method using Ser. rubidaea described in this report will facilitate the rapid identification of QSI-producing bacteria from marine environment. This study reveals new avenue for future environmental applications. This study also suggests that these algal epibiotic bacteria may play a role in the defensive mechanism for their host by producing QSI or QSI-like compounds to suppress the settlement of other competitive bacteria.     

Well-known quorum sensing inhibitors do not affect bacterial quorum sensing-regulated bean sprout spoilage

AIM: To investigate the potential of quorum sensing inhibitors (QSI) as food preservative agents in a food product, where bacterial spoilage is controlled by quorum sensing (QS). METHODS AND RESULTS: The effects of well-known QSI were tested on spoilage phenotypes and on QS-regulated genes of a bean sprout spoiling bacterial isolate (Pectobacterium A2JM) in laboratory substrates and in a bean sprout model system. The acylated homoserine lactones (AHL) analogues PenS-AHL and HepS-AHL decreased the specific protease activity of Pectobacterium A2JM in broth but did not reduce the expression of a QS-regulated secretion protein, and were without effect on soft rot of bean sprouts. The QSI ProS-AHL, furanone C-30, patulin, penicillic acid and 4-nitropyridine-N-oxide did not have any effect on protease activity, on gene expression or bean sprout appearance at nongrowth inhibitory concentrations. Extracts from garlic and bean sprouts induced the QS system of Pectobacterium in bean sprouts and a broth system, respectively. CONCLUSIONS: Among the several well-known QSI compounds, only PenS-AHL and HepS-AHL, inhibited QS-regulated protease activity of Pectobacterium A2JM in broth cultures, but had no effect on bean sprout spoilage. SIGNIFICANCE AND IMPACT OF THE STUDY: The QSI compounds must be selected in the specific system in which they are to function and they cannot easily be transferred from one QS system to another.     

An inhibitor of bacterial quorum sensing reduces mortalities caused by Vibriosis in rainbow trout (Oncorhynchus mykiss, Walbaum)

The fish pathogen Vibrio anguillarum produces quorum sensing signal molecules, N-acyl homoserine lactones (AHLs), which in several Gram-negative human and plant pathogenic bacteria regulate virulence factors. Expression of these factors can be blocked using specific quorum-sensing inhibitors (QSIs). The purpose of this study was to investigate the effect of a QSI, furanone C-30, on mortality of rainbow trout during challenge with V. anguillarum. Addition of 0.01 or 0.1 microM furanone C-30 to rainbow trout infected by cohabitation caused a significant reduction in accumulated mortality from 80-100% in challenge controls to 4-40% in treated groups. Furanone C-30 had no effect in an immersion challenge system, probably due to a very high water exchange and a rapid dilution of furanone C-30. Growth and survival of V. anguillarum were not affected by the concentrations of furanone C-30 used in the challenge experiments, thus avoiding selection for resistance. To elucidate the mechanism of disease control by furanone C-30, we determined its effect on the bacterial proteome, motility, and respiration. No effects were seen of furanone C-30 in any of these experiments. Although no cytotoxic effect on HeLa cells were observed, exposure to 1 microM (or higher) concentrations of furanone C-30 had detrimental effects on the rainbow trout. Our results indicate that QSIs can be used in non-antibiotic based control of fish diseases. However, they also underline the need for development of novel, less toxic QSI compounds and the need for understanding the exact mechanism(s) of action.     

微生物群体感应抑制剂及其在海洋生态中的应用

微生物具有结构多样性和功能多样性,其生态行为受多种信号因子的调节,其一便是群体感应信号(Quorum sensing,QS)。QS可作为菌群的通讯语言调节多种生物学功能,包括微生物被膜(Biofilm)的形成、毒力因子的表达、抗生素的分泌以及活性物质的生成等。相比之下,群体感应抑制剂(Quorum sensing inhibitor,QSI)的作用与QS相反,它能阻断QS信号的合成或传递、降低细菌致病性、干扰Biofilm的生成、阻断QS级联效应,因而被广泛应用于医药、农业和环境等领域。本文聚焦QSI,对其来源、特性、作用机制的最新进展进行总结,并对其在海洋生态领域上的应用进行综述,以期为QSI物质的开发和海洋生态资源的有效利用提供新思路。     

Quorum-sensing regulation in rhizobia and its role in symbiotic interactions with legumes

Legume-nodulating bacteria (rhizobia) usually produce N-acyl homoserine lactones, which regulate the induction of gene expression in a quorum-sensing (or population-density)-dependent manner. There is significant diversity in the types of quorum-sensing regulatory systems that are present in different rhizobia and no two independent isolates worked on in detail have the same complement of quorum-sensing genes. The genes regulated by quorum sensing appear to be rather diverse and many are associated with adaptive aspects of physiology that are probably important in the rhizosphere. It is evident that some aspects of rhizobial physiology related to the interaction between rhizobia and legumes are influenced by quorum sensing. However, it also appears that the legumes play an active role, both in terms of interfering with the rhizobial quorum-sensing systems and responding to the signalling molecules made by the bacteria. In this article, we review the diversity of quorum-sensing regulation in rhizobia and the potential role of legumes in influencing and responding to this signalling system.     

Disruption of bacterial quorum sensing by other organisms

Higher plants and algae produce compounds that mimic quorum sensing: signals used by bacteria to regulate the expression of many genes and behaviors. Similarly, various bacteria can stimulate, inhibit or inactivate quorum sensing in other bacteria. These discoveries offer new opportunities to manipulate bacterial quorum sensing in applications relevant to medicine, agriculture and the environment.     

Food as a source for quorum sensing inhibitors: iberin from horseradish revealed as a quorum sensing inhibitor of Pseudomonas aeruginosa

Foods with health-promoting effects beyond nutritional values have been gaining increasing research focus in recent years, although not much has been published on this subject in relation to bacterial infections. With respect to treatment, a novel antimicrobial strategy, which is expected to transcend problems with selective pressures for antibiotic resistance, is to interrupt bacterial communication, also known as quorum sensing (QS), by means of signal antagonists, the so-called QS inhibitors (QSIs). Furthermore, QSI agents offer a potential solution to the deficiencies associated with use of traditional antibiotics to treat infections caused by bacterial biofilms and multidrug-resistant bacteria. Several QSIs of natural origin have been identified, and in this study, several common food products and plants were extracted and screened for QSI activity in an attempt to isolate and characterize previously unknown QSI compounds active against the common opportunistic pathogen Pseudomonas aeruginosa. Several extracts displayed activity, but horseradish exhibited the highest activity. Chromatographic separation led to the isolation of a potent QSI compound that was identified by liquid chromatography-diode array detector-mass spectrometry (LC-DAD-MS) and nuclear magnetic resonance (NMR) spectroscopy as iberin-an isothiocyanate produced by many members of the Brassicaceae family. Real-time PCR (RT-PCR) and DNA microarray studies showed that iberin specifically blocks expression of QS-regulated genes in P. aeruginosa.