RESUMO
Cell-cell communication is critical for bacterial survival in natural habitats, in which miscellaneous regulatory networks are encompassed. However, elucidating the interaction networks of a microbial community has been hindered by the population complexity. This study reveals that γ-butyrolactone (GBL) molecules from Streptomyces species, the major antibiotic producers, can directly bind to the acyl-homoserine lactone (AHL) receptor of Chromobacterium violaceum and influence violacein production controlled by the quorum sensing (QS) system. Subsequently, the widespread responses of more Gram-negative bacterial AHL receptors to Gram-positive Streptomyces signaling molecules are unveiled. Based on the cross-talk between GBL and AHL signaling systems, combinatorial regulatory circuits (CRC) are designed and proved to be workable in Escherichia coli (E. coli). It is significant that the QS systems of Gram-positive and Gram-negative bacteria can be bridged via native Streptomyces signaling molecules. These findings pave a new path for unlocking the comprehensive cell-cell communications in microbial communities and facilitate the exploitation of innovative regulatory elements for synthetic biology.
Assuntos
4-Butirolactona/metabolismo , Acil-Butirolactonas/metabolismo , Proteínas de Bactérias/metabolismo , Bactérias Gram-Negativas/fisiologia , Bactérias Gram-Positivas/fisiologia , 4-Butirolactona/química , 4-Butirolactona/genética , 4-Butirolactona/farmacologia , Proteínas de Bactérias/genética , Chromobacterium/efeitos dos fármacos , Chromobacterium/genética , Chromobacterium/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Indóis/metabolismo , Interações Microbianas , Estrutura Molecular , Percepção de Quorum , Transdução de Sinais , Streptomyces/genética , Streptomyces/metabolismo , Biologia SintéticaRESUMO
Bacterial virulence factors facilitate host colonization and set the stage for the evolution of parasitic and mutualistic interactions. The Sodalis-allied clade of bacteria exhibit striking diversity in the range of both plant and animal feeding insects they inhabit, suggesting the appropriation of universal molecular mechanisms that facilitate establishment. Here, we report on the infection of the tsetse fly by free-living Sodalis praecaptivus, a close relative of many Sodalis-allied symbionts. Key genes involved in quorum sensing, including the homoserine lactone synthase (ypeI) and response regulators (yenR and ypeR) are integral for the benign colonization of S. praecaptivus. Mutants lacking ypeI, yenR and ypeR compromised tsetse survival as a consequence of their inability to repress virulence. Genes under quorum sensing, including homologs of the binary insecticidal toxin PirAB and a putative symbiosis-promoting factor CpmAJ, demonstrated negative and positive impacts, respectively, on tsetse survival. Taken together with results obtained from experiments involving weevils, this work shows that quorum sensing virulence suppression plays an integral role in facilitating the establishment of Sodalis-allied symbionts in diverse insect hosts. This knowledge contributes to the understanding of the early evolutionary steps involved in the formation of insect-bacterial symbiosis. Further, despite having no established history of interaction with tsetse, S. praecaptivus can infect reproductive tissues, enabling vertical transmission through adenotrophic viviparity within a single host generation. This creates an option for the use of S. praecaptivus in the biocontrol of insect disease vectors via paratransgenesis.
Assuntos
Percepção de Quorum/genética , Moscas Tsé-Tsé/genética , Fatores de Virulência/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/biossíntese , 4-Butirolactona/genética , Animais , Enterobacteriaceae/genética , Enterobacteriaceae/patogenicidade , Humanos , Insetos Vetores/genética , Insetos Vetores/microbiologia , Insetos/genética , Simbiose/genética , Moscas Tsé-Tsé/microbiologiaRESUMO
Pseudomonas aeruginosa, a versatile bacterium present in terrestrial and aquatic environments and a relevant opportunistic human pathogen, is largely known for the production of robust biofilms. The unique properties of these structures complicate biofilm eradication, because they make the biofilms very resistant to diverse antibacterial agents. Biofilm development and establishment is a complex process regulated by multiple regulatory genetic systems, among them is quorum sensing (QS), a mechanism employed by bacteria to regulate gene transcription in response to population density. In addition, environmental factors such as UVA radiation (400-315 nm) have been linked to biofilm formation. In this work, we further investigate the mechanism underlying the induction of biofilm formation by UVA, analysing the role of QS in this phenomenon. We demonstrate that UVA induces key genes of the Las and Rhl QS systems at the transcriptional level. We also report that pelA and pslA genes, which are essential for biofilm formation and whose transcription depends in part on QS, are significantly induced under UVA exposure. Finally, the results demonstrate that in a relA strain (impaired for ppGpp production), the UVA treatment does not induce biofilm formation or QS genes, suggesting that the increase of biofilm formation due to exposure to UVA in P. aeruginosa could rely on a ppGpp-dependent QS induction.
Assuntos
Biofilmes/efeitos da radiação , Pseudomonas aeruginosa/fisiologia , Percepção de Quorum/fisiologia , 4-Butirolactona/análogos & derivados , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Biofilmes/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica/efeitos da radiação , Genes Bacterianos/genética , Guanosina Tetrafosfato/genética , Guanosina Tetrafosfato/metabolismo , Mutação , Polissacarídeos Bacterianos/genética , Polissacarídeos Bacterianos/metabolismo , Pseudomonas aeruginosa/efeitos da radiação , Percepção de Quorum/genética , Percepção de Quorum/efeitos da radiação , Transcrição Gênica/efeitos da radiação , Raios UltravioletaRESUMO
The Acinetobacter baumanni J1 isolated from surface water of the Eastern Pacific Ocean, demonstrated significant algicidal activity on the algae Alexandrium tamarense. Interestingly, this strain showed the ability to produce an acyl-homoserine lactone (AHL) quorum sensing molecule. To better understand its AHL producing mechanism and its ecological functions, the genome of A.â¯baumanni strain J1 was completely sequenced. The genome contained a circular chromosome of 3,948,465â¯bp with an average GC content of 39.9â¯mol%. A total of 3707 protein coding genes, 41 tRNA genes and 16 rRNA genes were obtained. In silico genome annotation identified a LuxI putative gene located on contig 4. Subsequent thin-layer chromatography analysis indicated that C8-AHL could be produced by A.â¯baumanni J1, which confirmed the authenticity of the LuxI gene. Taken together, this work describes an algicidal bacterium that is capable of producing an AHL molecule, which may represent a valuable tool for developing microbial methods to control harmful algae.
Assuntos
4-Butirolactona/análogos & derivados , Acinetobacter baumannii/genética , Genoma Bacteriano , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Acinetobacter baumannii/metabolismo , Oceano Pacífico , Percepção de Quorum , Sequenciamento Completo do GenomaRESUMO
Quorum sensing (QS) in rhizobia regulates diverse processes determining the success and efficiency of association with the legume host. Despite the notable importance of QS as well as the well-known underlying variability in the genomic and metabolic components thereof, its study in rhizobia is largely restricted to few laboratory strains. In this work, QS phenomenon in the rhizobia nodulating pigeon pea- one of the most important legume crops of the global-south, is characterized. Using 16S rRNA and recombinaseA sequencing analysis, the selected QS-positive and host-beneficial isolates were identified to be taxonomically affiliated to the genus Ensifer. Their QS components, including homologues of QS genes, and the repertoire of N-acyl homoserine lactone (AHL) autoinducers were identified. Sequences of the QS homologues showed significant variabilities ranging from 10 to >20% with the known Ensifer sequences. Autoinducer profiling using LC-MS/MS revealed the production of long and short chain AHLs variably by the isolates, including 3-oxo-C12-homoserine lactone (3-O-C12-HSL) and 3-OH-C16-HSL as their first report in Rhizobiaceae. Motility and attachment- two of the most crucial traits for effective establishment on host roots were discovered to be QS dependent in in vitro analysis and the same was confirmed using expression analysis of their regulatory genes using qRT-PCR; both revealing a QS mediated repression of motility and promotion of attachment. This study highlights that Ensifer nodulating pigeon pea, although with significant variance in the anatomy of their QS components, regulate symbiotically crucial cell-processes via QS in a scheme that is conserved in multiple genera.
Assuntos
4-Butirolactona/análogos & derivados , Cajanus/microbiologia , Nodulação , Percepção de Quorum , Sinorhizobium , 4-Butirolactona/química , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Proteínas de Bactérias/genética , Cajanus/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica , Filogenia , Percepção de Quorum/genética , Percepção de Quorum/fisiologia , RNA Ribossômico 16S , Rhizobiaceae/classificação , Rhizobiaceae/isolamento & purificação , Rhizobiaceae/metabolismo , Sinorhizobium/isolamento & purificação , Sinorhizobium/metabolismo , SimbioseRESUMO
The interference of nonylphenol (NP) with humans and animals, especially in hormone systems, has been well-studied. There is rarely any record of its effect on bacteria, which dominate in various environments. In our study, we employed Pseudomonas aeruginosa PAO1 as a model microorganism and took its common lifestyle biofilm, mainly regulated by quorum sensing (QS), as a cut-in point to investigate the effect of NP (1, 5, 10â¯mgâ¯L-1) on bacteria. The results showed that more than 5â¯mgâ¯L-1 of NP did interfere with biofilm formation and affected bacterial QS. In detail, the LasI/R circuit, but not the RhlI/R circuit, was considerably obstructed. The decrease in lasI and lasR expression resulted in a significant reduction in N-3-oxo-dodecanoyl homoserine lactone (3OC12-HSL) signals and the downstream production of elastases. Docking results indicated the binding of NP with LasR protein, simulating the binding of 3OC12-HSL with LasR protein, which explained the obstruction of the LasIR circuit. We concluded that NP competed with 3OC12-HSL and blocked 3OC12-HSL binding with the LasR protein, resulting in a direct interference in bacterial biofilm formation. This is the first report of NP interference with bacterial signaling, which is not only helpful to understand the effect of NP on various ecosystems, but is also beneficial to enrich our knowledge of inter-kingdom communication.
Assuntos
Biofilmes/efeitos dos fármacos , Fenóis/toxicidade , Pseudomonas aeruginosa/efeitos dos fármacos , Percepção de Quorum/efeitos dos fármacos , 4-Butirolactona/análogos & derivados , 4-Butirolactona/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Pseudomonas aeruginosa/genéticaRESUMO
Lactone flavors with fruity, milky, coconut, and other aromas are widely used in the food and fragrance industries. Lactones are produced by chemical synthesis or by biotransformation of plant-sourced hydroxy fatty acids. We established a novel method to produce flavor lactones from abundant non-hydroxylated fatty acids using yeast cell factories. Oleaginous yeast Yarrowia lipolytica was engineered to perform hydroxylation of fatty acids and chain-shortening via ß-oxidation to preferentially twelve or ten carbons. The strains could produce γ-dodecalactone from oleic acid and δ-decalactone from linoleic acid. Through metabolic engineering, the titer was improved 4-fold, and the final strain produced 282â¯mg/L γ-dodecalactone in a fed-batch bioreactor. The study paves the way for the production of lactones by fermentation of abundant fatty feedstocks.
Assuntos
4-Butirolactona/análogos & derivados , Técnicas de Cultura Celular por Lotes , Ácido Linoleico/metabolismo , Ácido Oleico/metabolismo , Yarrowia , 4-Butirolactona/biossíntese , 4-Butirolactona/genética , Yarrowia/genética , Yarrowia/metabolismoRESUMO
MvaT and MvaU are global transcriptional regulators belonging to the H-NS family, and pyocyanin is an important virulence factor produced by Pseudomonas aeruginosa. mvaT mvaU double knockout mutant of P. aeruginosa PAO1 demonstrated pyocyanin abolishment in the previous study. Here, we further explored the mechanism. Two main directions were studied: pyocyanin biosynthesis pathway and QS system. The effect on the expression of the pyocyanin biosynthesis genes was evaluated by promoter strength determination and Real-Time PCR assay, and significant changes leading to low pyocyanin production were found. The effect on the QS system was studied by signal molecule quantification using LC-MS/MS and related gene expression measurements using Real-Time PCR. In mvaT mvaU double knockout, the production of 3-oxo-C12-HSL obviously increased, while those of C4-HSL and PQS obviously decreased, and the changes can be recovered by mvaT or mvaU complementation. The expressions of transcriptional activator genes binding with QS system signal molecules were all decreased, resulting in decreased formation of signal-transcriptional activator complexes. And the decreased expression of rhlR and pqsE also led to the lower expression of phzA1 and phzA2. Further exploration found that QS system downregulation may be related to QsrO, a QS system repressor, which was highly upregulated with mvaT mvaU double knockout. Hence, the synthesis of pyocyanin was suffocated and the biofilm formation ability was decreased. These results were also confirmed by transcriptome analysis, which demonstrated similar gene expression changes of the aforementioned genes together with decreased expression of other virulence factor genes regulated by QS system.
Assuntos
Proteínas de Bactérias/genética , Pseudomonas aeruginosa/genética , Piocianina/biossíntese , Transativadores/genética , Fatores de Virulência/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Animais , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Cromatografia Líquida , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Técnicas de Silenciamento de Genes , Homosserina/análogos & derivados , Homosserina/genética , Homosserina/metabolismo , Camundongos , Piocianina/genética , Percepção de Quorum/genética , Reação em Cadeia da Polimerase em Tempo Real , Espectrometria de Massas em TandemRESUMO
The link between quorum sensing in Vibrio campbellii and its virulence towards tiger grouper (Epinephelus fuscoguttatus) was investigated using V. campbellii wild type and quorum-sensing mutants with inactive quorum sensing or constitutively maximal quorum-sensing activity, and signal molecule synthase mutants. The results showed that wild-type V. campbellii is pathogenic to grouper larvae, causing more than 50% mortality after 4 days of challenge. Furthermore, the mortality of larvae challenged with the mutant with maximally active quorum sensing was significantly higher than that of larvae challenged with the wild type, whereas a higher survival was observed in the larvae challenged to the mutant with a completely inactive quorum-sensing system. Grouper larvae challenged with either the signal molecule synthase triple mutant, the harveyi autoinducer-1 (HAI-1) synthase mutant and the autoinducer-2 (AI-2) synthase mutant showed higher survival than larvae challenged with the wild type. In contrast, larvae challenged with the cholerae autoinducer-1 (CAI-1) synthase mutant showed high mortality. This indicates that HAI-1 and AI-2, but not CAI-1, are required for full virulence of V. campbellii towards grouper larvae. Our data suggest that quorum-sensing inhibition could be an effective strategy to control V. campbellii infections in tiger grouper.
Assuntos
Bass/microbiologia , Doenças dos Peixes/microbiologia , Percepção de Quorum , Vibrio/metabolismo , Vibrio/patogenicidade , 4-Butirolactona/análogos & derivados , 4-Butirolactona/genética , Animais , Homosserina/análogos & derivados , Homosserina/genética , Lactonas , Larva/microbiologia , Mutação , Vibrio/genética , VirulênciaRESUMO
Besides enzymatic conversions, many eukaryotic metabolic pathways also involve transport proteins that shuttle molecules between subcellular compartments, or into the extracellular space. Fungal itaconate production involves two such transport steps, involving an itaconate transport protein (Itp), and a mitochondrial tricarboxylate transporter (Mtt). The filamentous ascomycete Aspergillus terreus and the unicellular basidiomycete Ustilago maydis both produce itaconate, but do so via very different molecular pathways, and under very different cultivation conditions. In contrast, the transport proteins of these two strains are assumed to have a similar function. This study aims to investigate the roles of both the extracellular and mitochondrial transporters from these two organisms by expressing them in the corresponding U. maydis knockouts and monitoring the extracellular product concentrations. Both transporters from A. terreus complemented their corresponding U. maydis knockouts in mediating itaconate production. Surprisingly, complementation with At_MfsA from A. terreus led to a partial switch from itaconate to (S)-2-hydroxyparaconate secretion. Apparently, the export protein from A. terreus has a higher affinity for (S)-2-hydroxyparaconate than for itaconate, even though this species is classically regarded as an itaconate producer. Complementation with At_MttA increased itaconate production by 2.3-fold compared to complementation with Um_Mtt1, indicating that the mitochondrial carrier from A. terreus supports a higher metabolic flux of itaconic acid precursors than its U. maydis counterpart. The biochemical implications of these differences are discussed in the context of the biotechnological application in U. maydis and A. terreus for the production of itaconate and (S)-2-hydroxyparaconate.
Assuntos
Aspergillus/genética , Proteínas de Transporte/genética , Proteínas Fúngicas/genética , Ustilago/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/biossíntese , 4-Butirolactona/genética , Aspergillus/metabolismo , Proteínas de Transporte/metabolismo , Clonagem Molecular , Regulação Fúngica da Expressão Gênica , Técnicas de Inativação de Genes , Redes e Vias Metabólicas/genética , Mitocôndrias/genética , Succinatos/metabolismo , Ustilago/metabolismoRESUMO
Drug resistance is an obstacle to global malaria control, as evidenced by the recent emergence and rapid spread of delayed artemisinin (ART) clearance by mutant forms of the PfKelch13 protein in Southeast Asia. Identifying genetic determinants of ART resistance in African-derived parasites is important for surveillance and for understanding the mechanism of resistance. In this study, we carried out long-term in vitro selection of two recently isolated West African parasites (from Pikine and Thiès, Senegal) with increasing concentrations of dihydroartemisinin (DHA), the biologically active form of ART, over a 4-y period. We isolated two parasite clones, one from each original isolate, that exhibited enhanced survival to DHA in the ring-stage survival assay. Whole-genome sequence analysis identified 10 mutations in seven different genes. We chose to focus on the gene encoding PfCoronin, a member of the WD40-propeller domain protein family, because mutations in this gene occurred in both independent selections, and the protein shares the ß-propeller motif with PfKelch13 protein. For functional validation, when pfcoronin mutations were introduced into the parental parasites by CRISPR/Cas9-mediated gene editing, these mutations were sufficient to reduce ART susceptibility in the parental lines. The discovery of a second gene for ART resistance may yield insights into the molecular mechanisms of resistance. It also suggests that pfcoronin mutants could emerge as a nonkelch13 type of resistance to ART in natural settings.
Assuntos
4-Butirolactona/análogos & derivados , Artemisininas/farmacologia , Proteínas dos Microfilamentos/genética , Mutação/genética , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/genética , 4-Butirolactona/genética , Antimaláricos/farmacologia , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Resistência a Medicamentos/genética , Edição de Genes/métodos , Humanos , Malária Falciparum/tratamento farmacológico , Malária Falciparum/parasitologia , Repetições WD40/genéticaRESUMO
Butenolides are an emerging family of signaling molecules in Streptomyces. They control complex physiological traits, such as morphological differentiation and antibiotic production. However, how butenolides regulate these processes is poorly investigated because of obstacles in obtaining these signaling molecules. This study reports the identification of a butenolide-type signaling system for nikkomycin biosynthesis in Streptomyces ansochromogenes with distinct features. We identified a gene cluster, sab, consisting of three genes, sabAPD, for butenolide biosynthesis and two regulator genes, sabR1 and sabR2, and characterized three butenolides (SAB1, -2, and -3) by heterologous expression of sabAPD. sabA disruption abolished nikkomycin production, which could be restored by the addition of SABs or by deletion of sabR1 in ΔsabA. Electrophoretic mobility-shift assays and transcriptional analyses indicated that SabR1 indirectly represses the transcription of nikkomycin biosynthetic genes, but directly represses sabA and sabR1 In the presence of SABs, the SabR1 transcriptional regulator dissociated from its target genes, verifying that SabR1 is the cognate receptor of SABs. Genome-wide scanning with the conserved SabR1-binding sequence revealed another SabR1 target gene, cprC, whose transcription was strongly repressed by SabR1. Intriguingly, CprC positively regulated the pleiotropic regulatory gene adpA by binding to its promoter and, in turn, activated nikkomycin biosynthesis. This is the first report that butenolide-type signaling molecules and their cognate receptor SabR1 can regulate adpA via a newly identified activator, CprC, to control nikkomycin production. These findings pave the way for further studies seeking to unravel the regulatory mechanism and functions of the butenolide signaling system in Streptomyces.
Assuntos
4-Butirolactona/análogos & derivados , Aminoglicosídeos/metabolismo , Vias Biossintéticas , Streptomyces/metabolismo , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Aminoglicosídeos/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Família Multigênica , Streptomyces/genética , Ativação TranscricionalRESUMO
Many species of Proteobacteria produce acyl-homoserine lactone (AHL) compounds as quorum-sensing (QS) signals for cell density-dependent gene regulation. Most known AHL synthases, LuxI-type enzymes, produce fatty AHLs, and the fatty acid moiety is derived from an acyl-acyl carrier protein (ACP) intermediate in fatty acid biosynthesis. Recently, a class of LuxI homologs has been shown to use CoA-linked aromatic or amino acid substrates for AHL synthesis. By using an informatics approach, we found the CoA class of LuxI homologs exists primarily in α-Proteobacteria. The genome of Prosthecomicrobium hirschii, a dimorphic prosthecate bacterium, possesses a luxI-like AHL synthase gene that we predicted to encode a CoA-utilizing enzyme. We show the P. hirschii LuxI homolog catalyzes synthesis of phenylacetyl-homoserine lactone (PA-HSL). Our experiments show P. hirschii obtains phenylacetate from its environment and uses a CoA ligase to produce the phenylacetyl-CoA substrate for the LuxI homolog. By using an AHL degrading enzyme, we showed that PA-HSL controls aggregation, biofilm formation, and pigment production in P. hirschii These findings advance a limited understanding of the CoA-dependent AHL synthases. We describe how to identify putative members of the class, we describe a signal synthesized by using an environmental aromatic acid, and we identify phenotypes controlled by the aryl-HSL.
Assuntos
4-Butirolactona/análogos & derivados , Alphaproteobacteria/fisiologia , Proteínas de Bactérias , Biofilmes/crescimento & desenvolvimento , Proteínas de Transporte , Percepção de Quorum/fisiologia , Transdução de Sinais/fisiologia , 4-Butirolactona/biossíntese , 4-Butirolactona/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismoRESUMO
Microbial community with their plasticity follows a course of changes that allow adaptation and survival in a particular habitat. In this study perturbations in microbial flora dwelling in two reactors with phenol as a carbon source under the limiting nitrogen and phosphorus conditions were monitored for 3 months with alterations of dissolved oxygen (DO). With the time, the shift in diversity and abundance of bacteria were observed with simultaneous increase in biofilm-forming bacteria like Pseudomonas, Escherichia, etc. Functional level screening revealed that the abundance of core metabolic genes were not much altered, however, the regulated level of increase in quorum sensing genes (acyl-homoserine lactone), biofilm-forming genes, catalase and ferroxidase enzymes at high DO suggest the survival mechanism of the community. This study sheds light on survival route followed by the bacterial community with abiotic stress, such as an increase in DO.
Assuntos
Bactérias/genética , Estresse Oxidativo , Percepção de Quorum/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/genética , Bactérias/classificação , Bactérias/metabolismo , Fenômenos Fisiológicos Bacterianos , Biofilmes , Reatores Biológicos/microbiologia , Carbono/metabolismo , OxigênioRESUMO
Clostridium chauvoei causes fatal black quarter infection in cattle and buffaloes. The quorum sensing (QS) system, a bacterial cell to cell communication process, of the pathogen was characterized in the current study. The results indicated that C. chauvoei lacked luxS (autoinducer-2) based quorum sensing as detected by the sensor strain Vibrio harveyi BB170. This was supported by absence of luxS gene in C. chauvoei genome. However, the genomic analysis indicated the presence of agrBD system in all three genomes of C. chauvoei available at the NCBI database. The AgrD, which synthesizes QS messenger auto-inducing peptide, was a 44 amino acid protein which shared 59% identity and 75% similarity with AgrD of C. perfringens strain 13 and 56% identity (20% coverage) with Staphylococcus aureus N315. The functional cysteine amino acid was conserved in all the strains. The genomic organisation further suggests the presence of diguanylate cyclase, a gene responsible for synthesis of secondary messenger cyclic di-GMP, at 3' immediate downstream of agrD gene. The real time expression analysis for agrD gene indicated that expression was better at 37⯰C (1.9-3.7 fold increase) compared to a higher temperature of 40⯰C. However, stable expression was observed at different growth stages (log and early stationary phase) with 0.8-1.4 fold changes in expression pattern. The results indicate the presence of a constitutively expressed agrBD quorum sensing system in C. chauvoei.
Assuntos
Proteínas de Bactérias/metabolismo , Doenças dos Bovinos/microbiologia , Infecções por Clostridium/veterinária , Clostridium chauvoei/fisiologia , Percepção de Quorum , 4-Butirolactona/análogos & derivados , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Animais , Proteínas de Bactérias/genética , Bovinos , Infecções por Clostridium/microbiologia , Clostridium chauvoei/genética , Clostridium chauvoei/crescimento & desenvolvimento , Regulação Bacteriana da Expressão GênicaRESUMO
Covering: 2000 to 2018 The antimicrobial activity of many of their natural products has brought prominence to the Streptomycetaceae, a family of Gram-positive bacteria that inhabit both soil and aquatic sediments. In the natural environment, antimicrobial compounds are likely to limit the growth of competitors, thereby offering a selective advantage to the producer, in particular when nutrients become limited and the developmental programme leading to spores commences. The study of the control of this secondary metabolism continues to offer insights into its integration with a complex lifecycle that takes multiple cues from the environment and primary metabolism. Such information can then be harnessed to devise laboratory screening conditions to discover compounds with new or improved clinical value. Here we provide an update of the review we published in NPR in 2011. Besides providing the essential background, we focus on recent developments in our understanding of the underlying regulatory networks, ecological triggers of natural product biosynthesis, contributions from comparative genomics and approaches to awaken the biosynthesis of otherwise silent or cryptic natural products. In addition, we highlight recent discoveries on the control of antibiotic production in other Actinobacteria, which have gained considerable attention since the start of the genomics revolution. New technologies that have the potential to produce a step change in our understanding of the regulation of secondary metabolism are also described.
Assuntos
Actinobacteria/genética , Actinobacteria/metabolismo , Antibacterianos/metabolismo , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Antraquinonas/metabolismo , Antibacterianos/biossíntese , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Carbono/metabolismo , Regulação Bacteriana da Expressão Gênica , Microbiologia Industrial/métodos , Família Multigênica , Nitrogênio , Metabolismo Secundário , Estreptomicina/biossíntese , Estreptomicina/metabolismoRESUMO
Quorum sensing is a widespread form of cell-to-cell communication, which is based on the production of signaling molecules known as autoinducers (AIs). The first group contains highly species-specific N-acyl homoserine lactones (N-AHLs), generally known as AI-1, which are produced by AHL synthase. The second group, possessing the characteristic structure of a furanone ring, are known as AI-2. The enzyme responsible for their production is S-ribosylhomocysteine lyase (LuxS). In Campylobacter jejuni, AI-2 and LuxS play a role in many important processes, including biofilm formation, stress response, motility, expression of virulence factors, and colonization. However, neither the receptor protein nor the exact structure of the AI-2 molecule have been identified to date. Similarly, little is known about the possible existence of AHL-synthase producing AI-1 and its impact on gene expression. Recently, an analogue of homoserine lactone, called cjA, was isolated from a cell-free supernatant of C. jejuni strain 81-176 and from the food isolate c11. The molecule cjA particularly impacted the expression of virulence factors and biofilm formation. This review summarizes the role of AI-2 and cjA in the context of biofilm formation, motility, stress responses, and expression of virulence factors.
Assuntos
4-Butirolactona/análogos & derivados , Campylobacter jejuni/metabolismo , Regulação Bacteriana da Expressão Gênica , Homosserina/análogos & derivados , Lactonas/metabolismo , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Campylobacter jejuni/genética , Campylobacter jejuni/patogenicidade , Homosserina/genética , Homosserina/metabolismo , Percepção de Quorum , Especificidade da Espécie , Estresse Fisiológico/fisiologia , Fatores de Virulência/metabolismoRESUMO
Human coronin 3 is involved in many types of cancers, but the underlying molecular mechanisms require further elucidation. The present study demonstrated that coronin 3 is significantly upregulated in clinical primary hepatocellular carcinoma (HCC) samples by reverse transcriptionquantitative polymerase chain reaction (RTqPCR) and immunohistochemical staining. Subsequently, proteins that were regulated by coronin 3 in both coronin 3 overexpressing or knocked down HepG2 cells were analyzed by label free mass spectrometry; overall, 249 proteins were identified to be closely regulated by coronin 3, and those coronin 3 regulated proteins were enriched in cellular, physiological and metabolism processes. By further indepth pathway analysis, it was demonstrated that those proteins were involved into 94 different pathways. Finally, the expression levels of glucose6phosphatase catalytic subunit 3 (G6PC3) were confirmed to be negatively regulated by coronin 3, as determined by RTqPCR and western blotting. In conclusion, these results indicated that coronin3 is significantly dysregulated in HCC tumor tissues, and may exert its function via regulating G6PC3 expression. These results provide valuable information for further study of coronin 3mediated signaling pathways, and implicate coronin 3 as a potential therapeutic target for HCC.
Assuntos
4-Butirolactona/análogos & derivados , Glucose-6-Fosfatase/metabolismo , Espectrometria de Massas , 4-Butirolactona/genética , 4-Butirolactona/metabolismo , Adulto , Idoso , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Ontologia Genética , Glucose-6-Fosfatase/genética , Células Hep G2 , Humanos , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Coloração e Rotulagem , Proteínas rac1 de Ligação ao GTP/metabolismoRESUMO
Sodalis praecaptivus is a close relative and putative environmental progenitor of the widely distributed, insect-associated, Sodalis-allied symbionts. Here we show that mutant strains of S. praecaptivus that lack genetic components of a quorum-sensing (QS) apparatus have a rapid and potent killing phenotype following microinjection into an insect host. Transcriptomic and genetic analyses indicate that insect killing occurs as a consequence of virulence factors, including insecticidal toxins and enzymes that degrade the insect integument, which are normally repressed by QS at high infection densities. This method of regulation suggests that virulence factors are only utilized in early infection to initiate the insect-bacterial association. Once bacteria reach sufficient density in host tissues, the QS circuit represses expression of these harmful genes, facilitating a long-lasting and benign association. We discuss the implications of the functionality of this QS system in the context of establishment and evolution of mutualistic relationships involving these bacteria.