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Unravelling osmotololerance in Escherichia coli using a pseudosexual model system

ABSTRACT: Transcriptional analysis of mutants evolved for tolerance to excess sodium chloride (up to 0.75 M)

ORGANISM(S): Escherichia coli BW25113 Escherichia coli Escherichia coli O157:H7 str. EDL933 Escherichia coli O157:H7 str. Sakai Escherichia coli CFT073 Escherichia coli str. K-12 substr. MG1655

PROVIDER: GSE51611 | GEO | 2014/04/16

SECONDARY ACCESSION(S): PRJNA224098

REPOSITORIES: GEO

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Unravelling osmotololerance in Escherichia coli using a pseudosexual model system

Project description:Transcriptional analysis of mutants evolved for tolerance to excess sodium chloride (up to 0.75 M) Mutant compared to parental reference during sustained exponential growth in 0.55 M NaCl, two biological replicates per sample, reference is unevolved parent strain for each mutant

2014-04-16 | E-GEOD-51611 | biostudies-arrayexpress

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Proteomics Analysis of Evolved Escherichia coli Populations from Cyclic Antibiotic Treatment

Project description:When the survivors of antibiotic treatment (persisters) are repeatedly regrown and retreated with the same antibiotic for several cycles, the new population will soon adapt to the treatment condition and become tolerant to the drug. Here, we did evolution experiments on Escherichia coli populations by treating it with daily high concentration of different antibiotics (ampicillin, ciprofloxacin and apramycin) approximating clinical dosage, during the rapid growth-exponential phase. After a few cycles, we observed that the evolved populations exhibit extremely high tolerance to the drug, which are achieved by single point mutations in one of several genes. Interestingly, treatment with different antibiotics led to the selection of different mutants despite the shared persistence phenotype. Here, we applied spectral counting-based quantitative proteomics to study the proteome profile of the evolved E. coli populations from different cyclic antibiotic treatments.

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Evolution combined with genomic study elucidates genetic bases of isobutanol tolerance in Escherichia coli

Project description:BACKGROUND:Isobutanol is a promising next generation biofuel with demonstrated high yield microbial production, but the toxicity of this molecule reduces fermentation volumetric productivity and final titers. Organic solvent tolerance is a complex, multigenic phenotype that has been recalcitrant to rational engineering approaches. We apply experimental evolution followed by genome resequencing and a gene expression study to elucidate genetic bases on adaptation to exogenous isobutanol stress. RESULTS:The adaptations acquired in our evolved lineages exhibit antagonistic pleiotropy between minimal and rich medium, and appear to be specific to the effects of longer chain alcohols. By examining genotypic adaptation in multiple independent lineages, we find evidence of parallel evolution in hfq, mdh, acrAB, gatYZABCD, and rph genes. Many isobutanol tolerant lineages show reduced rpoS activity, perhaps related to mutations in hfq or acrAB. Consistent with the complex, multigenic nature of solvent tolerance, we observe adaptations in a diversity of cellular processes. Many adaptations appear to involve epistasis between different mutations, implying a rugged fitness landscape for isobutanol tolerance. We observe a trend of evolution targeting post-transcriptional regulation and high centrality nodes of biochemical networks. Collectively, the genotypic adaptations we observe suggest mechanisms of adaptation to isobutanol stress based on remodelling the cell envelope and surprisingly, stress response attenuation. CONCLUSIONS:We have discovered a set of genotypic adaptations that confer increased tolerance to exogenous isobutanol stress. Our results are immediately useful to efforts to engineer more isobutanol tolerant host strains of E. coli for isobutanol production. We suggest that rpoS and post-transcriptional regulators, such as hfq, RNA helicases, and sRNAs may be interesting mutagenesis targets for futurue global phenotype engineering.

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