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Evidence of a role for CutRS and actinorhodin in the secretion stress response in Streptomyces coelicolor M145

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Streptomyces Coelicolor A3(2)

TISSUE(S): Cell Culture, Cell Line Cell

SUBMITTER: Gerhard Saalbach

LAB HEAD: Matt Hutchings

PROVIDER: PXD040579 | Pride | 2023-06-16

REPOSITORIES: Pride

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Proteomic analysis of Streptomyces coelicolor vesicles

Project description:Streptomyces coelicolor is a model organism for the study of Streptomyces, a genus of Gram-positive bacteria that undergoes a complex life cycle and produces an enormous repertoire of bioactive metabolites and extracellular enzymes. This study investigated the production and characterization of membrane vesicles (MVs) in liquid cultures of S. coelicolor M145 from a structural and biochemical point of view using microscopic, physical and -omics analyzes. Two main populations of MVs, F3 and F4 MVs, with different size and cargo were isolated and purified. S. coelicolor MV cargo is complex and contains many “messages” such as proteins and metabolites. A total of 166 proteins involved in cell metabolism and differentiation, molecular processing and transport was identified in MVs. In particular, a subset of these proteins was protected from the degradation after proteinase K treatment, indicating their localization inside the vesicles. Vesicle proteome includes many stress response proteins which also play an important role in S. coelicolor morpho- physiological differentiation. Moreover, MVs packaged with an array of metabolites such as antibiotics, vitamins, amino acids and components of carbon metabolism. The analysis of S. coelicolor MV cargo will provide informations to elucidate their biogenesis and functions

2021-11-02 | PXD024085 | Pride

Differential impact of phosphate availability on primary and specialized metabolism of Streptomyces lividans and Streptomyces coelicolor.

Project description:In order to define the impact of phosphate (Pi) availability on cellular metabolism the project aimed to perform a comparative analysis of the proteomes of two Streptomyces strains with different abilities to produce antibiotics, S. coelicolor and S. lividans as well as of the pptA mutant of S. lividans, grown low (1mM) and high (5mM) phosphate (Pi) availability conditions. Interestingly, in contrast to most Streptomyces species, S. coelicolor produces more antibiotics in Pi proficiency than in Pi limitation, S. lividans does not produce antibiotics in any Pi conditions and the pptA mutant produces antibiotics only in Pi limitation. This in-depth proteomic comparison of three Streptomyces strains (S. coelicolor, S. lividans wt and pptA mutant), in different growth conditions (time and Pi concentration in the medium) was performed on four biological replicates. Protein abundance changes were determined using two label-free mass spectrometry based-quantification methods: spectral count (SC) and MS1 ion intensities named XIC (for eXtracted Ion Current). Our proteomic data reveal for the first time, the impact of Pi availability on the abundance of approximately 4000 proteins of these Streptomyces strains with different abilities to produce antibiotics. The most striking feature differentiating these strains was the much higher abundance of enzymes of the respiratory chain in both phosphate conditions in S. coelicolor compared to the S. lividans strains.

2022-02-04 | PXD029263 | Pride

Transcription profiling by array of Streptomyces coelicolor actinorhodin-producing wild type andrecombinant strains

Project description:Genome-wide expression analysis of 6 batch cultivations of actinorhodin-producing wild type and recombinant strain of Streptomyces coelicolor

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RNA-seq of Streptomyces coelicolor and Streptomyces davawensis including a dodecin gene deletion variant in response to plumbagin treatment

Project description:Genes encoding dodecin proteins are present in almost 20% of archaeal and in more than 50% of bacterial genomes. Archaeal dodecins bind riboflavin (vitamin B2), are thought to play a role in flavin homeostasis and possibly also help to protect cells from radical or oxygenic stress. Bacterial dodecins were found to bind riboflavin-5’-phosphate (also called flavin mononucleotide or FMN) and coenzyme A, but their physiological function remained unknown. In this study, we set out to investigate the relevance of dodecins for flavin metabolism and oxidative stress management in the phylogenetically related bacteria Streptomyces coelicolor and Streptomyces davawensis. Therfore we have treated Streptomyces coelicolor wildtype, Streptomyces davawensis wildytype and Streptomyces davawensis dodecin deletion strain with plumbagin, a compound, which induces oxidative stress in exponential and stationary growth phase and analysed the transcriptome via RNA-seq (Illumina TruSeq stranded mRNA libraries sequenced on Illumina HiSeq rapid mode 2 x 70nt PE).

2018-04-07 | E-MTAB-6633 | biostudies-arrayexpress

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Project description:Comparative genomic hybridization analysis of Streptomyces coelicolor A3(2) versus Streptomyces lividans 66 and Streptomyces lividans TK24 using high density 105,000 x 60-mer ink-jet in situ synthesized arrays.

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Morphological differentiation and secondary metabolism responses of Streptomyces coelicolor A3(2) to simulated microgravity based on comparative transcriptomics.

Project description:In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to simulated microgravity condition by clinorotation, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturerM-bM-^@M-^Ys recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each. Cells of Streptomyces coelicolor A3(2) wild type strain were harvested from the colonies grown in JCM42 agar medium at 28M-BM-0C for 96h (4d) under simulated microgravity and 1g control conditions. Total RNA was extracted to conduct the microarray hybridization experiments. Three independent experiments were performed in which ASM_1A, ASM_2A, ASM_3A were the test group samples and ANG_1A, ANG_2A, ANG_3A were the control group samples. Differentially expressed genes were selected with PM-oM-<M-^\0.05, FCM-bM-^IM-%2.0 by T-test methods using the independent three sets data.

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