Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

ROS homeostasis and pimaricin production in Streptomyces natalensis ATCC 27448

ABSTRACT: The aim of this work was to unveil the molecular mechanisms by which Streptomyces respond to a ROS intracellular imbalance and the effect of such response on the biosynthesis of secondary metabolites. The study was focused on the industrial actinomycete S. natalensis ATCC 27448 producer of the polyene pimaricin - an antifungal agent widely used in the food industry and promising for antiviral activity and stimulation of immune response. Two-color microarray with common reference. The transcriptomes of S. natalensis ATCC 27448 (wild-type), S. natalensis CAM.02 (DsodF) and S. natalensis CAM.04 (DahpCD) were compared. Two time points were included: late exponential (T1) and early stationay (T2) phase. Biological triplicates were performed for each strain/time point. Genomic DNA of S. natalensis ATCC 27448 was used as common reference.

ORGANISM(S): Streptomyces natalensis ATCC 27448

SUBMITTER: Tiago Beites

PROVIDER: E-GEOD-46537 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Genome-wide analysis of the regulation of pimaricin production in Streptomyces natalensis by reactive oxygen species.

Beites Tiago T Rodríguez-García Antonio A Santos-Beneit Fernando F Moradas-Ferreira Pedro P Aparicio Jesús F JF Mendes Marta V MV

Applied microbiology and biotechnology 20140111 5

To investigate the molecular mechanisms that interplay between oxygen metabolism and secondary metabolism in Streptomyces natalensis, we compared the transcriptomes of the strains CAM.02 (ΔsodF), pimaricin under-producer phenotype, and CAM.04 (ΔahpCD), pimaricin over-producer phenotype, with that of the wild type at late exponential and stationary growth phases. Microarray data interpretation was supported by characterization of the mutant strains regarding enzymatic activities, phosphate uptake ...[more]

PMID: 24413916

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Project description:To accelerate previous RNA structure probing approaches, which focus on analyzing one RNA sequence at a time, we have developed FragSeq, a high-throughput RNA structure probing method that uses high-throughput RNA sequencing to identify single-stranded RNA (ssRNA) regions from fragments generated by nuclease P1, which is specific for single-stranded nucleic acids. In the accompanying study, we show that we can accurately and simultaneously map ssRNA regions in multiple non-coding RNAs with known structure in experiments probing the entire mouse nuclear transcriptome. We carried out probing in two cell types to assess reproducibility. We also identified and experimentally validated structured regions in ncRNAs never previously probed. We examined mouse nuclear RNA from two cell types: undifferentiated embryonic stem cells (UNDIFF) and cells differentiated into neural precursors (D5NP). For each cell type, nuclear RNA was purified and deproteinized, denatured, and refolded in vitro, from which we prepared three barcoded samples: "nuclease" (RNA partially digested with P1 ssRNA-specific nuclease, yielding 5'-PO4/3'-OH end chemistry at each cleavage site), "control" (control for "nuclease" sample to idenfity endogenous 5'-PO4/3'-OH), and "PNK" (same as "control" but followed by a polynucleotide kinase treatment to convert 5'-OH/3'-cyclic-phosphate ends to clonable 5'-PO4/3'-OH ends). Resulting RNA fragments were cloned using the SOLiD Small RNA Expression Kit (SREK) protocol, which ligates linkers only to 5'-PO4/3'-OH containing RNA, enriching for clones of products resulting from P1 cleavage in "nuclease" sample and selecting against random degradation. Two cell types, three treatments each, thus resulted in six barcoded samples total (barcodes 01, 02, 04, 05, 07, 08). Four other barcoded samples were prepared for separate experiments not used in our study (barcodes 03, 06, 09, 10), so their preparation is not described here. The total run of ten barcodes was done on the ABI SOLiD3 platform and a custom algorithm (FragSeq v0.0.1) was used to compute "cutting scores" (as described in our paper) that show ssRNA regions in hundreds of ncRNAs.

Dataset Information

ROS homeostasis and pimaricin production in Streptomyces natalensis ATCC 27448

Publications

Genome-wide analysis of the regulation of pimaricin production in Streptomyces natalensis by reactive oxygen species.

Similar Datasets

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