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TraDIS-Xpress finds mechanisms of action and synergies of a novel lipid IVA biosynthesis inhibitor

ABSTRACT: TraDIS-Xpress finds mechanisms of action and synergies of a novel lipid IVA biosynthesis inhibitor

PROVIDER: PRJEB65052 | ENA |

REPOSITORIES: ENA

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TraDIS-Xpress finds mechanisms of action and synergies of a novel lipid IVA biosynthesis inhibitor

Project description:We used TraDIS-Xpress to determine the mechanism of action of a novel antimicrobial compound. We found that it inhibits lipid IVA biosynthesis in both Escherichia coli and Salmonella enterica serovar Typhimurium. We also were able to determine mechanisms of synergy with colistin, through ATP biosynthesis and the BasSR signalling system.

2023-08-31 | E-MTAB-13250 | biostudies-arrayexpress

TraDIS-Xpress analysis of colistin stress responses in Escherichia coli

Project description:In past, resistance mechanisms have been identified by analysis of resistant isolates or defined mutants. Recently, high-throughput transposon mutagenesis coupled with sequencing (TraDIS-Xpress) is another approach proving useful for elucidating the roles of genes involved in the overall cellular response to a particular stress. In this study, we used TraDIS-Xpress to determine the role played by genes following exposure to colistin stress. Approximately 10^7 cells from the mutant library were inoculated into LB broth at a range of doubling concentrations of colistin ( 0.25 x MIC, 0.5 x MIC, 1 x MIC, 2 X MIC). Experiments were performed with no induction, or with induction using 0.2 or 1 mM of Isopropyl β-D-1-thiogalactopyranoside (IPTG). All experiments were performed in duplicate.

2022-06-09 | E-MTAB-11807 | biostudies-arrayexpress

TraDIS-Xpress to determine E. coli response to Piperacillin and Tazobactam separately and in combination

Project description:Found how E. coli responds to the commonly prescribed antibiotic-inhibitor combination piperacillin and tazobactam, separately and in combination. TraDIS-Xpress was performed following the protocol outlined by Yasir et al 2020 Genome Research. The results suggest tazobactam triggers the activity of multidrug efflux systems that have been previously seen to confer clinical resistance to multiple classes of antibiotics.

2023-08-31 | E-MTAB-13226 | biostudies-arrayexpress

TraDIS-Xpress analysis of gene functions in Escherichia coli

Project description:TraDIS-Xpress analysis of gene functions in Escherichia coli

| PRJEB53418 | ENA

TraDIS-Xpress analysis of colistin stress responses in Escherichia coli

Project description:TraDIS-Xpress analysis of colistin stress responses in Escherichia coli

| PRJEB53417 | ENA

Investigating Biofilm Formation Mechanisms in Salmonella on Fresh Produce Using TraDIS-Xpress

Project description:Investigating Biofilm Formation Mechanisms in Salmonella on Fresh Produce Using TraDIS-Xpress

| PRJEB67878 | ENA

TraDIS-Xpress to determine E. coli response to Piperacillin and Tazobactam separately and in combination

Project description:TraDIS-Xpress to determine E. coli response to Piperacillin and Tazobactam separately and in combination

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TraDIS-Xpress: a high-resolution whole-genome assay identifies novel mechanisms of triclosan action and resistance.

Project description:Understanding the genetic basis for a phenotype is a central goal in biological research. Much has been learnt about bacterial genomes by creating large mutant libraries and looking for conditionally important genes. However, current genome-wide methods are largely unable to assay essential genes which are not amenable to disruption. To overcome this limitation, we developed a new version of "TraDIS" (transposon directed insertion-site sequencing) that we term "TraDIS-Xpress" that combines an inducible promoter into the transposon cassette. This allows controlled overexpression and repression of all genes owing to saturation of inserts adjacent to all open reading frames as well as conventional inactivation. We applied TraDIS-Xpress to identify responses to the biocide triclosan across a range of concentrations. Triclosan is endemic in modern life, but there is uncertainty about its mode of action with a concentration-dependent switch from bacteriostatic to bactericidal action unexplained. Our results show a concentration-dependent response to triclosan with different genes important in survival between static and cidal exposures. These genes include those previously reported to have a role in triclosan resistance as well as a new set of genes, including essential genes. Novel genes identified as being sensitive to triclosan exposure include those involved in barrier function, small molecule uptake, and integrity of transcription and translation. We anticipate the approach we show here, by allowing comparisons across multiple experimental conditions of TraDIS data, and including essential genes, will be a starting point for future work examining how different drug conditions impact bacterial survival mechanisms.

| S-EPMC7050523 | biostudies-literature

Staphylococcus aureus strain:SA0831-iva | isolate:SA0831-iva

Project description:Staphylococcus aureus strain:SA0831-iva | isolate:SA0831-iva Genome sequencing

| PRJNA1176700 | ENA

Triclosan TraDIS sequencing

Project description:Triclosan TraDIS sequencing

| PRJEB29311 | ENA