Virulence and bacteriocin of Escherichia coli ST131 and other E. coli lineages.
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PROVIDER: PRJEB34226 | ENA |
REPOSITORIES: ENA
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Project description:Escherichia coli ST131 is a clinical challenge due to its multidrug resistant profile and successful global spread. They are often associated with complicated infections, particularly urinary tract infections (UTIs). Bacteriocins play an important role to outcompete other microorganisms present in the human gut. Here, we characterized bacteriocin-encoding plasmids found in ST131 isolates of patients suffering from a UTI using both short- and long-read sequencing. Colicins Ia, Ib and E1, and microcin V, were identified among plasmids that also contained resistance and virulence genes. To investigate if the potential transmission range of the colicin E1 plasmid is influenced by the presence of a resistance gene, we constructed a strain containing a plasmid which had both the colicin E1 and blaCMY-2 genes. No difference in transmission range was found between transformant and wild-type strains. However, a statistically significantly difference was found in adhesion and invasion ability. Bacteriocin-producing isolates from both ST131 and non-ST131 lineages were able to inhibit the growth of other E. coli isolates, including other ST131. In summary, plasmids harboring bacteriocins give additional advantages for highly virulent and resistant ST131 isolates, improving the ability of these isolates to compete with other microbiota for a niche and thereby increasing the risk of infection.
| S-EPMC6920910 | biostudies-literature
Project description:Escherichia coli sequence type 131 (ST131) is a globally dominant multidrug resistant clone associated with urinary tract and bloodstream infections. Most ST131 strains exhibit resistance to multiple antibiotics and cause infections associated with limited treatment options. The largest sub-clonal ST131 lineage is resistant to fluoroquinolones, contains the type 1 fimbriae fimH30 allele and expresses an H4 flagella antigen. Flagella are motility organelles that contribute to UPEC colonisation of the upper urinary tract. In this study, we examined the specific role of H4 flagella in ST131 motility and interaction with host epithelial and immune cells. We show that the majority of H4-positive ST131 strains are motile and are enriched for flagella expression during static pellicle growth. We also tested the role of H4 flagella in ST131 through the construction of specific mutants, over-expression strains and isogenic mutants that expressed alternative H1 and H7 flagellar subtypes. Overall, our results revealed that H4, H1 and H7 flagella possess conserved phenotypes with regards to motility, epithelial cell adhesion, invasion and uptake by macrophages. In contrast, H4 flagella trigger enhanced induction of the anti-inflammatory cytokine IL-10 compared to H1 and H7 flagella, a property that may contribute to ST131 fitness in the urinary tract.
| S-EPMC4637896 | biostudies-other
Project description:Secretion of recombinant proteins to the culture medium after transport to the periplasm of gram-negative bacteria often facilitates downstream-processing. It could lead to more biological active and correct folded proteins and low contamination with host proteins, thereby reducing the costs of the production process. However, Escherichia coli K12 secrets only little amount and few substrates naturally. This is one of the major drawbacks for the applicability of this strain in the biotechnology industry, when large amounts of biological active proteins are desirable. Therefore, different attempts were made to enhance the level of secreted proteins. Coexpression of bacteriocin release proteins makes the outer membrane permeable for proteins. A detailed insight into the complex underlying regulatory mechanism and metabolic changes when such release proteins are expressed could be very useful for successful optimisation strategies. The identification of potential optimisation targets can be achieved by DNA-microarray-technology, as proven in many cases before. In this work DNA-microarrays were used for the identification of differentially expressed genes of an inducible E.coli secretion strain expressing reporter proteins bacterial alkaline phosphatase, PhoA and beta-lactamase, Bla and their release to the culture medium by coexpression of the BRP of CloDF13. Based on the data of whole genome experiments and on the different databases genes which are regulated after induction of BRP expression are identified and discussed. BRP activity was found to cause a substantial cellular response and considerable changes in global gene expression. Additionally, performed cluster-analysis using k-means algorithm identified clusters containing promising candidates for new optimisation strategies aiming for an enhanced protein secretion.
2009-08-01 | GSE16946 | GEO
Project description:Proteomic Characterization of Bacteriocin-Producing Enterococcus faecium Strains from foodstuffs
2023-03-11 | PXD037241 | Pride
Project description:Escherichia coli ST131 plasmids
| PRJNA494636 | ENA
Project description:Secretion of recombinant proteins to the culture medium after transport to the periplasm of gram-negative bacteria often facilitates downstream-processing. It could lead to more biological active and correct folded proteins and low contamination with host proteins, thereby reducing the costs of the production process. However, Escherichia coli K12 secrets only little amount and few substrates naturally. This is one of the major drawbacks for the applicability of this strain in the biotechnology industry, when large amounts of biological active proteins are desirable. Therefore, different attempts were made to enhance the level of secreted proteins. Coexpression of bacteriocin release proteins makes the outer membrane permeable for proteins. A detailed insight into the complex underlying regulatory mechanism and metabolic changes when such release proteins are expressed could be very useful for successful optimisation strategies. The identification of potential optimisation targets can be achieved by DNA-microarray-technology, as proven in many cases before. In this work DNA-microarrays were used for the identification of differentially expressed genes of an inducible E.coli secretion strain expressing reporter proteins bacterial alkaline phosphatase, PhoA and beta-lactamase, Bla and their release to the culture medium by coexpression of the BRP of CloDF13. Based on the data of whole genome experiments and on the different databases genes which are regulated after induction of BRP expression are identified and discussed. BRP activity was found to cause a substantial cellular response and considerable changes in global gene expression. Additionally, performed cluster-analysis using k-means algorithm identified clusters containing promising candidates for new optimisation strategies aiming for an enhanced protein secretion. time series of BRP Induction, every sample was labeled with Cy3 and Cy5 (Dye Swap) to compensate dye-specific effects.
2009-08-01 | E-GEOD-16946 | biostudies-arrayexpress
Project description:Background: Treponema denticola is strongly associated with the development of periodontal disease. Both synergistic and antagonistic effects are observed among bacterial species in the process of biofilm formation. Bacteriocin-related genes have not yet been fully characterized in periodontopathic bacteria. The aim of this study was to detect and characterize bacteriocin-associated proteins in T. denticola. Methods: The whole genome sequence of T. denticola ATCC 35405 was screened with a Streptococcus mutans bacteriocin immunity protein (ImmA/Bip) sequence. The prevalence of homologous genes in T. denticola strains was then investigated by Southern blotting. Expression of the genes was evaluated by qRT-PCR. Results: In the genome sequence of T. denticola, an amino acid sequence coded by open reading frame TDE_0719 showed 26% identity with the S. mutans ImmA. Furthermore, two protein sequences coded by TDE_0425 and TDE_2431 in T. denticola ATCC 35405 showed ~40% identity with that coded by TDE0719. Therefore, TDE_0425, TDE_0719, and TDE_2431 were designated as tepA1, A2, and A3, respectively. Open reading frames showing similarity to the HlyD family of secretion proteins were detected downstream of tepA1, A2, and A3. They were designated as tepB1, B2, and B3, respectively. A gene harboring a bacteriocin-like signal sequence was detected upstream of tepA1. The prevalence of tepA1 and A2 differed among Treponema species. Susceptibility to chloramphenicol and ofloxiacin was slightly decreased in a tepA2 mutant while that to kanamycin was increased. Expression of tepA3-B3 was increased in the tepA2 mutant. Conclusion: These results indicate that T. denticola ATCC 35405 has three potential bacteriocin export proteins and that the presence of these genes differs among the Treponema strains. These proteins may be involved in resistance to chloramphenicol.
2017-01-01 | GSE83445 | GEO
Project description:A popular strategy for enhancing the antibacterial properties of probiotic bacteria is to retrofit them with the ability to overproduce heterologous bacteriocins. This is often achieved from strong, non-native promoters. How the dysregulated overproduction of heterologous bacteriocins affects the fitness and antibacterial efficacy of the retrofitted probiotic bacteria is often overlooked. We conferred the prototypical probiotic Escherichia coli strain Nissle (EcN) the ability to produce different amounts of the bacteriocin microcin C (McC). Expression of the bacteriocin synthesis genes was driven from the native promoter (Pmcc-WT), or from promoters manipulated to be stronger (Pmcc-High) and weaker (Pmcc-Low) than the WT, in a plasmid-based system. Pmcc-Low and Pmcc-High retained their native regulation. A strain harbouring a non-functional promoter (Pmcc-Mut) produces no McC and was used as a control. Each strain was grown to early stationary phase, when production of McC starts, in Luria-Bertani broth at 37 degrees. The RNA was isolated and the effects of different levels of production of McC on the transcriptome of EcN was examined by RNA-Seq.
2023-06-30 | E-MTAB-13078 | biostudies-arrayexpress
Project description:This is the first report of a bacteriocin being produced by lactobacillus acidophilus ATCC 4356. For protein identification, the ~8-kDa peptide band was removed from the acrylamide gel and digested in-gel by trypsin, and the resulting peptide fragments were extracted and analyzed by LC-MS/MS analysis. The MS data were processed using Thermo Proteome Discoverer software (v2.2) with the SEQUEST search engine. Three peptides, VAHCASQIGR (amino acids 23-32), GSAACVSYLTR (amino acids 69-79), GSAACVSYLTRHRHH (amino acids 69-83) were identified as tryptic fragments on the basis of LC-MS/MS.This is the first report of a bacteriocin being produced by lactobacillus acidophilus ATCC 4356. For protein identification, the ~8-kDa peptide band was removed from the acrylamide gel and digested in-gel by trypsin, and the resulting peptide fragments were extracted and analyzed by LC-MS/MS analysis. The MS data were processed using Thermo Proteome Discoverer software (v2.2) with the SEQUEST search engine. Three peptides, VAHCASQIGR (amino acids 23-32), GSAACVSYLTR (amino acids 69-79), GSAACVSYLTRHRHH (amino acids 69-83) were identified as tryptic fragments on the basis of LC-MS/MS.
2020-03-24 | PXD011556 | Pride
Project description:The induction of genetic competence is a strategy used by bacteria to increase their genetic repertoire under stressful environmental conditions. Recently, Streptococcus pneumoniae has been shown to co-ordinate the uptake of transforming DNA with fratricide via increased expression of the peptide pheromone responsible for competence induction. Here, we document that environmental stress-induced expression of the peptide pheromone competence-stimulating peptide (CSP) in the oral pathogen Streptococcus mutans. We showed that CSP is involved in the stress response and determined the CSP-induced regulon in S. mutans by microarray analysis. Contrary to pneumococcus, S. mutans responds to increased concentrations of CSP by cell lysis in only a fraction of the population. We have focused on the mechanism of cell lysis and have identified a novel bacteriocin as the ‘death effector’. Most importantly, we showed that this bacteriocin causes cell death via a novel mechanism of action: intracellular action against self. We have also identified the cognate bacteriocin immunity protein, which resides in a separate unlinked genetic locus to allow its differential regulation. The role of the lytic response in S. mutans competence is also discussed. Together, these findings reveal a novel autolytic pathway in S. mutans which may be involved in the dissemination of fitness-enhancing genes in the oral biofilm.
2010-05-29 | GSE22019 | GEO