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Global effects of pathway-specific regulator CpkO and CpkN deletion in Streptomyces coelicolor A3(2)

ABSTRACT: Relative quantification of protein abundances of S. coelicolor A3(2) parent strain M145 and deletion mutants ∆cpkO and ∆cpkN. This study shows novel insights into the mechanism of coelimycin synthesis regulation and the network of interactions between cpk cluster regulatory proteins and the biosythetic genes of other secondary metabolite clusters.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Streptomyces Coelicolor A3(2)

TISSUE(S): Prokaryotic Cell

SUBMITTER: Aaron Millan Oropeza

LAB HEAD: Celine Henry

PROVIDER: PXD012672 | Pride | 2021-03-29

REPOSITORIES: Pride

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	20181023_04_P193_A.mzXML	Mzxml
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	20181023_06_P196_A.mzXML	Mzxml
	20181023_06_P196_A.xml	Xml
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Relative quantification from chemostats cultures of Saccharomyces cerevisiae CEN.PK113-7D, Kluyveromyces marxianus CBS 6556 and Yarrowia lipolytica W29

Project description:Relative quantification of protein abundances of three yeast strains (Saccharomyces cerevisiae CEN.PK113-7D, Kluyveromyces marxianus CBS6556 and Yarrowia lipolytica W29) cultivate in chemostats under different conditions. The conditions for Saccharomyces cerevisiae CEN.PK113-7D are: - Standard condition – 30°C, pH 5.5 - High temperature - 36°C, pH 5.5 - Low pH - 30°C, pH 3.5 - Osmotic stress – 30°C, pH 5.5, 1M KCl The conditions for Kluyveromyces marxianus CBS6556 are: - Standard condition – 30°C, pH 5.5 - High temperature - 40°C, pH 5.5 - Low pH - 30°C, pH 3.5 - Osmotic stress – 30°C, pH 5.5, 0.6 M KCl The conditions for Yarrowia lipolytica W29 are: - Standard condition - 28°C, pH 5.5 - High temperature - 32°C, pH 5.5 - Low pH - 28°C, pH 3.5 This study is part of the OMICS data generation of CHASSY project (European Union’s Horizon 2020 grant agreement No 720824).

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RNA-Sequenzing of Streptomyces coelicolor A3(2) (M1152) in timeseries

Project description:SYSTERACT: Systematic Rebuilding of Actinomycetes for Natural Product Formation For several decades antibiotics have saved millions of lives, but their overuse makes them less effective due to increase in bacterial resistance. Because of this major clinical and public health problem, there is an urgent need for new effective antimicrobials. The ERASysAPP project SYSTERACT aims to further develop, the model actinobacterium Streptomyces coelicolor into improved microbial cell factories to heterologously produce diverse bioactive compounds in amounts needed for structural and functional evaluation. Unprecedented systems biology understanding of S. coelicolor is being combined with morphology engineering and improved (de-)regulation and precursor supply to accelerate bioactive compound discovery efforts. By that means, we aim to generate a stepwise improved 'Superhost' for the production of antibiotics in which metabolic bottlenecks and regulatory restriction are greatly mitigated. The optimized strains will be tested concerning their applicability for an improved production of commercially relevant antibiotics and the expression of novel bioactive gene clusters identified in new actinomycete strains and environmental metagenomes. So far two strains, M145 and M1152, have been cultivated for time-resolved 'omics sampling, and a larger number of additional strains are on the list for similar experiments. High quality RNAseq-based transcriptome data have been generated and processed. M145 is the wildtype strain in S. coelicolor (as used in STREAM, see also GSE18489), 3 biol. replicas and M1152 lacks four major biosynthetic gene clusters, undecylprodigine (RED), calcium-dependent antibiotic (CDA), coelimycin (CPK) and actinorhodin (ACT). Contributors: A. Wentzel, W. Wohlleben, G. van Wezel, D van Dissel, O. Wolkenhauer, E. Kerkhoven, N. Spidsoe, K. Nieselt and the SYSTERACT consortium

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Project description:The chromosomes of several widely used laboratory derivatives of Streptomyces coelicolor A3(2) were found to have 1.06 Mb inverted repeat sequences at their termini (i.e. long-terminal inverted repeats; L-TIRs), which are 50 times the length of the 22 kb TIRs of the sequenced S. coelicolor strain M145. The L-TIRs include 1005 annotated genes and increase the overall chromosome size to 9.7 Mb. The 1.06 Mb L-TIRs are the longest reported thus far for an actinomycete, and are proposed to represent the chromosomal state of the original soil isolate of S. coelicolor A3(2). S. coelicolor A3(2), M600 and J1501 possess L-TIRs, whereas approximately half the examined early mutants of A3(2) generated by ultraviolet (UV) or X-ray mutagenesis have truncated their TIRs to the 22 kb length. UV radiation was found to stimulate L-TIR truncation. Two copies of a transposase gene (SCO0020) flank 1.04 Mb of DNA in the right L-TIR, and recombination between them appears to generate strains containing short TIRs. This TIR reduction mechanism may represent a general strategy by which transposable elements can modulate the structure of chromosome ends. The presence of L-TIRs in certain S. coelicolor strains represents a major chromosomal alteration in strains previously thought to be genetically similar.

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