Project description:Bacteria in the genus Streptomyces are soil dwelling oligotrophs and important producers of secondary metabolites. Previously we showed that global mRNA expression was subject to a series of metabolic and regulatory switches during the life time of a fermentor batch culture of S. coelicolor M145. Here we analyse the proteome from eight time points from the same fermentor culture and, as phosphate availability is an important regulator of secondary metabolite production, compare this to the proteome of a similar time course from an S. coelicolor mutant, INB201 (ΔphoP), defective in the control of phosphate utilisation. The proteomes provide a detailed view of enzymes involved in central carbon and nitrogen metabolism. Trends in protein expression over the time courses were deduced from a protein abundance index which also revealed the importance of stress pathway proteins in both cultures. As expected the ΔphoP mutant was deficient in expression of PhoP-dependent genes and several putatively compensatory metabolic and regulatory pathways for phosphate scavenging were detected. Notably there is a succession of switches that co-ordinately induce the production of enzymes for five different secondary metabolite biosynthesis pathways over the course of the batch cultures and these were not confined to the stationary phase.
Project description:Bacteria in the genus Streptomyces are soil dwelling oligotrophs and important producers of secondary metabolites. Previously we showed that global mRNA expression was subject to a series of metabolic and regulatory switches during the life time of a fermentor batch culture of S. coelicolor M145. Here we analyse the proteome from eight time points from the same fermentor culture and, as phosphate availability is an important regulator of secondary metabolite production, compare this to the proteome of a similar time course from an S. coelicolor mutant, INB201 (M-NM-^TphoP), defective in the control of phosphate utilisation. The proteomes provide a detailed view of enzymes involved in central carbon and nitrogen metabolism. Trends in protein expression over the time courses were deduced from a protein abundance index which also revealed the importance of stress pathway proteins in both cultures. As expected the M-NM-^TphoP mutant was deficient in expression of PhoP-dependent genes and several putatively compensatory metabolic and regulatory pathways for phosphate scavenging were detected. Notably there is a succession of switches that co-ordinately induce the production of enzymes for five different secondary metabolite biosynthesis pathways over the course of the batch cultures and these were not confined to the stationary phase. 36 samples, no replicates; one hour resolution from 23-36h and 41-48h; half hour resolution from 36-41h; two hour resolution 48-60h; sample missing for 34 h
Project description:During the lifetime of a fermenter culture, the soil bacterium S. coelicolor undergoes a major metabolic switch from exponential growth to antibiotic production. We have studied gene expression patterns during this switch, using a specifically designed Affymetrix GeneChip and a high-resolution time-series of fermenter-grown samples. This time series was conducted using medium leading to glutamate depletion and the cultivation conditions as published in Nieselt et al. BMC Genomics 2010, performed with the Streptomyces coelicolor wild type strain M145E.
Project description:This work was carried out to elucidate the proteins that are regulated by the two-component system CutRS in Streptomyces coelicolor M145 and how this response changes in the presence of glucose. A comparison of the whole cell proteomes of Streptomyces coelicolor M145 WT and Streptomyces coelicolor M145 ∆cutRS on both DNA (no glucose) and DNAD (with glucose) was made.
Project description:Alam2010 - Genome-scale metabolic network of
Streptomyces coelicolor
This model is described in the article:
Metabolic modeling and
analysis of the metabolic switch in Streptomyces
coelicolor.
Alam MT, Merlo ME, STREAM
Consortium, Hodgson DA, Wellington EM, Takano E, Breitling
R.
BMC Genomics 2010; 11: 202
Abstract:
BACKGROUND: The transition from exponential to stationary
phase in Streptomyces coelicolor is accompanied by a major
metabolic switch and results in a strong activation of
secondary metabolism. Here we have explored the underlying
reorganization of the metabolome by combining computational
predictions based on constraint-based modeling and detailed
transcriptomics time course observations. RESULTS: We
reconstructed the stoichiometric matrix of S. coelicolor,
including the major antibiotic biosynthesis pathways, and
performed flux balance analysis to predict flux changes that
occur when the cell switches from biomass to antibiotic
production. We defined the model input based on observed
fermenter culture data and used a dynamically varying objective
function to represent the metabolic switch. The predicted
fluxes of many genes show highly significant correlation to the
time series of the corresponding gene expression data.
Individual mispredictions identify novel links between
antibiotic production and primary metabolism. CONCLUSION: Our
results show the usefulness of constraint-based modeling for
providing a detailed interpretation of time course gene
expression data.
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Project description:We identified genome-wide binding regions of NdgR in Streptomyces coelicolor using chromatin immunoprecipitation sequencing (ChIP-seq). We constructed 6×myc-tagged NdgR strain using homologous recombination with myc-tagging vector. Analysis of the sequencing data aligned to Streptomyces coelicolor genome database (NC_003888).
Project description:S. coelicolor was grown in low-phosphate concentration R5 liquid medium which facilitated the transition to a phosphate starvation induced stationary phase and the subsequent induction of the PhoP regulon. The genome wide binding locations of PhoP during phosphate starvation were determined by chromatin-immunopreciptiation followed by hybridisation to high density S. coelicolor DNA microarrays. The use of both wild-type S. coelicolor and phoP-null mutant strains allowed the identification of specific PhoP binding locations.
Project description:To identify unique gene expression in cAMP supplemented Streptomyces coelicolor M1146 strain. The genes with different gene expression might be key genes to understand the effects of cAMP supplementation on the transcriptome of Streptomyces coelicolor M1146.
Project description:To identify unique gene expression in cAMP supplemented Streptomyces coelicolor M145 strain. The genes with different gene expression might be key genes to understand the effects of cAMP supplementation on the transcriptome of Streptomyces coelicolor M145.
Project description:Genome-wide expression analysis of 6 batch cultivations of actinorhodin-producing wild type and recombinant strain of Streptomyces coelicolor