Project description:This dataset contains the gene expression signature in triplicates of Escherichia coli BW25113 growing exponentially on eight different environments: minimal medium supplemented with 5 g/L of Glucose, Galactose, Glycerol, Gluconate, Fructose, Pyruvate, Succinate or Acetate as the sole carbon source.
Project description:To understand the gene response during the glucose to acetate diauxic transition, we grew E. coli in minimal media with acetate and a small amount of glucose. Cells were collected and RNA was purified at different time points during the growth transition, including pre-shift (growth on glucose), 5 minutes, 15 minutes, 60 minutes and 120 minutes after glucose run-out, and steady state growth on acetate (post-shift).
Project description:This experiment was performed to challenge E. coli response to acetate when its growth rate flux is reduced by 40% with 100 mM methyl α-D-glucopyranoside. Control without inhibition in the exact same conditions was already published on ArrayExpress database (accession number E-MTAB-9086) and could be used for comparison purpose. Here, E. coli BW25113 strain was grown in M9 medium complemented with 15 mM glucose, 100 mM αMG, and 0, 10, 50 or 100 mM sodium acetate. The cells were grown to mid-exponential phase in shake flasks at 37 °C and 200 rpm, in 50 mL of medium before proceeding to RNA extraction and subsequent microarray analysis. Three to four replicates (with two to three biological replicates) were analyzed for each condition. We thank all the INSA-Toulouse students that participated in this project: Lola Blayac, Romane Ducloux, Benjamin Jung, Oliver Larousse, Leyre Sarrias, Arno Bruel, Alexis Charbinat, Justine Dumas-Perdriau, Solène Frapard, Sophie Germain, Nicolas Papadopoulos, Marine Rodeghiero & Auriane Thomas.
Project description:Specific growth rate dependent gene expression changes of Escherichia coli K12 MG1655 were determined by microarray and real time PCR analyses. The bacteria were cultivated on glucose limited minimal medium using the accelerostat method (A-stat), where starting from steady state conditions in a chemostat culture, dilution rate is constantly increased. At specific growth rate (μ) 0.47 h-1, E. coli had focused its metabolism to glucose utilization by down-regulation of alternative substrate transporters expression compared to μ = 0.3 h-1. It was found that acetic acid accumulation began at μ = 0.34 ± 0.01 h-1 and two acetate synthesis pathways (phosphotransacetylase-acetate kinase (pta-ackA) and pyruvate oxidase (poxB)) contributed to the synthesis at the beginning of overflow metabolism, i.e. onset of acetate excretion. On the other hand, poxB, pta and ackA expression patterns suggest that pyruvate oxidase may be the only enzyme synthesizing acetate at μ = 0.47 h-1. Down-regulation of acs-yjcH-actP operon, the resulting loss of glucose and acetate co-utilization between specific growth rates 0.3 h-1 – 0.42 h-1 and acetic acid accumulation from μ = 0.34 ± 0.01 h-1 allows one to surmise that the acetate utilization operon expression might play an important role in overflow metabolism.
Project description:Escherichia coli K-12 BW25113 were cultured in the LB and M63 media. Exponetially growing populations were collected for RNAseq. The culture medium-induced transcriptional changes were analyzed.
Project description:Specific growth rate dependent gene expression changes of Escherichia coli K12 MG1655 were determined by microarray and real time PCR analyses. The bacteria were cultivated on glucose limited minimal medium using the accelerostat method (A-stat), where starting from steady state conditions in a chemostat culture, dilution rate is constantly increased. At specific growth rate (μ) 0.47 h-1, E. coli had focused its metabolism to glucose utilization by down-regulation of alternative substrate transporters expression compared to μ = 0.3 h-1. It was found that acetic acid accumulation began at μ = 0.34 ± 0.01 h-1 and two acetate synthesis pathways (phosphotransacetylase-acetate kinase (pta-ackA) and pyruvate oxidase (poxB)) contributed to the synthesis at the beginning of overflow metabolism, i.e. onset of acetate excretion. On the other hand, poxB, pta and ackA expression patterns suggest that pyruvate oxidase may be the only enzyme synthesizing acetate at μ = 0.47 h-1. Down-regulation of acs-yjcH-actP operon, the resulting loss of glucose and acetate co-utilization between specific growth rates 0.3 h-1 â 0.42 h-1 and acetic acid accumulation from μ = 0.34 ± 0.01 h-1 allows one to surmise that the acetate utilization operon expression might play an important role in overflow metabolism. DNA microarray analysis was performed from three A-stat cultivations, for which one has a technical replicate.
Project description:Bacteria have to continuously adjust to nutrient fluctuations from favorable to less favorable conditions and carbon starvation. The glucose-acetate transition followed by carbon starvation is representative of such carbon fluctuations observed by E. coli in many environments. Regulation of gene expression through fine-tuning of mRNA pools constitutes one of the regulation levels required for such a metabolic adaptation. It results from both mRNA transcription and degradation controls. However, the contribution of transcript stability regulation in gene expression is poorly characterized. Using combined transcriptome and mRNA decay analyses, we investigated (i) how transcript stability changes in E. coli during the glucose-acetate-starvation transition and (ii) if these changes contribute to gene expression changes. Our work highlights that transcript stability increases along carbon depletion. Most of the stabilization occurs at glucose-acetate transition when glucose is exhausted, then stabilized mRNAs remain stable during acetate consumption and carbon starvation. Meanwhile, expression of most genes is downregulated and we observed three time less gene expression upregulation. Using control analysis theory on 375 genes, we show that most of gene expression regulation is driven by changes in transcription. Although mRNA stabilization is not the controlling phenomenon, it contributes to the emphasis or attenuation of transcription regulation. However, upregulation of 18 genes (33% of our studied upregulated set) is mainly governed by transcript stabilization. Because these genes are associated with response to nutrient changes and stress, this illustrates the importance of post-transcriptional regulations in bacterial response to nutrient starvation.
Project description:Bacteria have to continuously adjust to nutrient fluctuations from favorable to less favorable conditions and carbon starvation. The glucose-acetate transition followed by carbon starvation is representative of such carbon fluctuations observed by E. coli in many environments. Regulation of gene expression through fine-tuning of mRNA pools constitutes one of the regulation levels required for such a metabolic adaptation. It results from both mRNA transcription and degradation controls. However, the contribution of transcript stability regulation in gene expression is poorly characterized. Using combined transcriptome and mRNA decay analyses, we investigated (i) how transcript stability changes in E. coli during the glucose-acetate-starvation transition and (ii) if these changes contribute to gene expression changes. Our work highlights that transcript stability increases along carbon depletion. Most of the stabilization occurs at glucose-acetate transition when glucose is exhausted, then stabilized mRNAs remain stable during acetate consumption and carbon starvation. Meanwhile, expression of most genes is downregulated and we observed three time less gene expression upregulation. Using control analysis theory on 375 genes, we show that most of gene expression regulation is driven by changes in transcription. Although mRNA stabilization is not the controlling phenomenon, it contributes to the emphasis or attenuation of transcription regulation. However, upregulation of 18 genes (33% of our studied upregulated set) is mainly governed by transcript stabilization. Because these genes are associated with response to nutrient changes and stress, this illustrates the importance of post-transcriptional regulations in bacterial response to nutrient starvation.