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TraDIS-Xpress analysis of colistin stress responses in Escherichia coli

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

PROVIDER: PRJEB53417 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR9830850.fastq.gz	Fastqsanger.gz
	ERR9830860.fastq.gz	Fastqsanger.gz
	ERR9830870.fastq.gz	Fastqsanger.gz
	ERR9830851.fastq.gz	Fastqsanger.gz
	ERR9830861.fastq.gz	Fastqsanger.gz

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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 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 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

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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

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

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Proteomic changes of Klebsiella pneumoniae in response to colistin treatment and crrB mutation-mediated colistin resistance

Project description:Polymyxins are increasingly used as the critical last-resort therapeutic options for multidrug-resistant gram-negative bacteria. Unfortunately, polymyxin resistance has increased gradually for the last few years. Although studies on mechanisms of polymyxin are expanding, system-wide analyses of the underlying mechanism for polymyxin resistance and stress response are still lacking. To understand how Klebsiella pneumoniae adapt to colistin (polymyxin E) pressure, we carried out proteomic analysis of Klebsiella pneumoniae strain cultured with different concentrations of colistin. Our results showed that the proteomic responses to colistin treatment in Klebsiella pneumoniae involving several pathways, including (i) gluconeogenesis and TCA cycle; (ii) arginine biosynthesis; (iii) porphyrin and chlorophyll metabolism; and (iv) enterobactin biosynthesis. Interestingly, decreased abundance of class A β-lactamases including TEM, SHV-11, SHV-4 were observed in cells treated with colistin. Moreover, we also present comprehensive proteome atlases of paired polymyxin-susceptible and -resistant Klebsiella pneumoniae strains. The polymyxin-resistant strain Ci, a mutant of Klebsiella pneumoniae ATCC BAA 2146, showed missense mutation in crrB. The crrB mutant Ci, which displayed lipid A modification with 4-amino-4-deoxy-L-arabinose (L-Ara4N) and palmitoylation, showed striking increases of CrrAB, PmrAB, PhoPQ, ArnBCADT and PagP. We hypothesize that crrB mutations induce elevated expression of the arnBCADTEF operon and pagP via PmrAB and PhoPQ. Moreover, multidrug efflux pump KexD, which was induced by crrB mutation, also contributed to colistin resistance. Overall, our results demonstrated proteomic responses to colistin treatment and the mechanism of CrrB-mediate colistin resistance, which may further offer valuable information to manage polymyxin resistance.

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Project description:We found that the antibiotic colistin acts synergistically with antifungals of the echinocandin class (e.g. aminocandin) on C. albicans cells. In order to elucidate the mode of action of colistin in fungi we performed microarray analysis of samples treated with only aminocandin (0.00125µg/ml) or treated with aminocandin (0.00125µg/ml) and colistin (5µg/ml). We compared: (A). untreated cells to cells treated with aminocandin only; (B). cells treated with aminocandin to cells treated with aminocandin and colistin (which is the focus of this experiment). By comparing those datasets it should be possible to identify genes differentially expressed in response to aminocandin and in response to both drugs. And subsequently to be able to interpret where in the cell colistin acts. (See related experiment in ArrayExpress: E-MEXP-3437 for comparison between untreated cells vs cells treated with aminocandin only.)

2012-03-03 | E-MEXP-3438 | biostudies-arrayexpress

Colistin resistant A. baumannii: genomic and transcriptomic traits acquired under colistin therapy

Project description:Comparative Genomic and Transcriptomic Characterization of Colistin-Resistant Acinetobacter baumannii

2019-01-23 | GSE109951 | GEO