Project description:Streptomyces sp. M7 has demonstrated ability to remove lindane from culture media and soils. In this study, we used MS-based label-free quantitative proteomic to understand lindane degradation and its metabolic context in Streptomyces sp. M7. We identified the proteins involved in the up-stream degradation pathway. Our results demonstrated that mineralization of lindane is feasible since proteins from an unusual down-stream degradation pathway were also identified. Degradative steps were supported by an active catabolism that supplied energy and reducing equivalents in the form of NADPH. This is the first study in which degradation steps of an organochlorine compound and metabolic context are elucidate in a biotechnological genus as Streptomyces. These results serve as basement to study other degradative actinobacteria and to improve the degradation processes of Streptomyces sp. M7.
Project description:Chip-seq to identify the HrdB (SCO5820) binding sites and regulated genes in Streptomyces coelicolor. The HrdB gene was tagged on genome by HA (Human Influenza hemagglutinin derived) epitope (TAC CCG TAC GAT GTG CCG GAT TAC GCG). We have 3 replicates with HrdB tagged by HA and 2 replicates with wild-type strain as controls. Chip-seq experiment was conducted using antibody against HA tag in vegetative growth phase (see protocols).
Project description:This study compared the genome of Streptomyces rimosus rimosus against that of Streptomyces coelicolor. It also compared 4 strains with changes in oxytetracycline production and derived from G7, the type strain, against G7. Keywords: Comparative genomic hybridization
Project description:BldC is a transcriptional regulator essential for morphological development Streptomyces venezuelae. Although bldC deletion strain is unable to produce aerial hyphae, electron microscopy reveals that almost all of the colony biomass is in the form of spores rather than undifferentiated vegetative hyphae. This ChIP-chip experiment was carried out to determine the binding sites, and thence the regulon, of BldC in Streptomyces venezuelae. Cy3(IP):Cy5(Total) signal ratios in the wild type were compared to those in a bldC knockout strain.
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.
Project description:Chip-seq experiment used to identify the binding sites of alternative σ factor σE in Streptomyces coelicolor. We access the binding sites with and without EtOH stress condition (see methods). To capture the binding of σE, the σE gene was tagged on genome by HA (Human Influenza hemagglutinin derived) epitope (TAC CCA TAC GAC GTC CCA GAC TAC GCT) on its C-terminus, wild type strain without tagged σE served as negative control.
Project description:We performed ribosome profiling which is the deep-sequencing of mRNA fragments protected by translating ribosome for two Streptomyces species through different growth phases to provide the translatome data
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).
Project description:Living organisms are exposed on a daily basis to widespread mixtures of toxic compounds. Mixtures pose a major problem in the assessment of health effects because they often generate substance-specific effects that cannot be attributed to a single mechanism. Two compounds often found together in the environment are the heavy metal chromium and the polycyclic aromatic hydrocarbon benzo[a]pyrene (B[a]P). We have examined how long-term exposure to a low concentration of Cr(VI) affects the transcriptional response to B[a]P, a second toxicant with an unrelated mechanism of action. Growth of mouse hepatoma cells for 20 passages in medium with 0.1 or 0.5 uM Cr(VI) increases DNA damage and apoptosis and decreases clonogenic ability. These cells also show transcriptome changes indicative of increased expression of DNA damage response and repair genes. In them, B[a]P activates cancer progression pathways, unlike cells never exposed to Cr(VI), where B[a]P activates mostly xenobiotic metabolism pathways. Cells grown in Cr(VI) for 20 passages and then cultured for an additional 5 passages in the absence of Cr(VI) recover from some but not all the chromium effects. They show B[a]P-dependent transcriptome changes strongly weighted towards xenobiotic metabolism, similar to those in B[a]P-treated cells that had no previous Cr(VI) exposure, but retain a high level of Cr(VI)-induced DNA damage and silence the expression of DNA damage and cancer progression genes. We conclude that the combined effect of these two toxicants appears to be neither synergistic nor cumulative, generating a toxic/adaptive condition that cannot be predicted from the effect of each toxicant alone. Mouse Hepa-1 cells treated with Cr, Bap, Cr+Bap and untreated were profiled using RNAseq in duplicates.