Project description:We report the mRNA and small RNA transcriptomes of Streptomyces coelicolor, Streptomyces avermitilis, and Streptomyces venezuelae. We identified dozens of new conserved sRNAs and antisense RNAs, including a prominent group of antisense RNAs termed ‘cutoRNAs’ that result from overlap of the 3′ ends of convergently transcribed mRNAs. In addition, we observed abundant unique ncRNAs, including many within secondary metabolic gene clusters.
Project description:To determine the influence of BldM and WhiI on the binding of each other to their respective binding sites in the genome of Streptomyces venezuelae, ChIP-Seq experiments were carried out using polyclonal antibodies against the wild type proteins as well as anti-FLAG antibodies against FLAG-tagged proteins. Null mutants of Streptomyces venezuelae lacking WhiI or BldM were used as controls. The wild tpe strain was used as control in experiments using the anti-FLAG antibodies.
Project description:We discovered the bacteria Streptomyces venezuelae can display a new form of growth termed exploration, and we used NGS to compare transcriptome profiles of cells demonstrating exploration versus static cells (not demonstrating exploration)
Project description:We isolated and sequenced mRNA from Streptomyces venezuelae grown on solid medium that promotes exploratory behaviour in this bacterial species. The data was analyzed using DeSeq2 to identify genes that undergo changes in expression over time.
Project description:The whiH gene is required for the differentiation of aerial hyphae into spores in Streptomyces species. It is a predicted member of the GntR family of transcription factors and has been shown to bind specifically to a sequence in its own promoter. This ChIP-Seq experiment was carried out to determine all the binding sites whiH binds to in the genome of Streptomyces venezuelae. A whiH deletion strain was made and a FLAG tagged whiH protein was expressed in it from a genome-integrated plasmid. Then anti-FLAG antibodies were used for chromatin immunoprecipitation followed by high throughput sequencing. The wild type Streptomyces venezuelae strain (ATCC 10712) was used as a negative control. For both the FLAG-WhiH strain and the WT strain, non-immunoprecipitated (total) DNA was also sequenced to arrive at a background enrichment which could be subtracted from the enrichment in the immunoprecipated sample.
Project description:We studied the binding of SMC protein to Streptomyces venezuelae chromosome during development of aerial hyphae (14 hour of growth) using SMC-FLAG protein. Additionally investigated the role of ParB protein in SMC DNA binding using parB deletion strain.
Project description:The WhiG sigma factor gene is required for spore formation is Streptomyces venezuelae. It is similar to the FliA sigma factor of E. coli. WhiG deletion strains are able to make aerial hyphae but are defective in the spore maturation. This ChIP-Seq experiment was carried out to determine all the binding sites WhiG binds to in the genome of Streptomyces venezuelae. Anti-WhiG polyclonal antibodies were used for ChIP-Seq of the wild type (WT) strain after 34 hours of growth in shaken cultures. A WhiG deletion strain was made and anti-WhiG antibodies were used for ChIP-Seq in the deletion strain after 34 hours of growth in shaken cultures. This was used as the negative control and ChIP-Seq peak positions in this were disregarded in the WT.
Project description:To examine the effects of cyclic-di-GMP on DNA binding by BldD in vivo, we manipulated the levels of c-di-GMP in Streptomyces venezuelae and monitored the effect on BldD binding to its target promoters in vivo by ChIP-seq, using a polyclonal BldD antibody. The degree of BldD binding was assayed at a single time point in wild-type (wt) S. venezuelae and the wt overexpressing either the diguanylate cyclase CdgB or the phosphodiesterase YhjH. A bldD null mutant was used a negative control.
Project description:The BldC gene is required for the formation of aerial hyphae Streptomyces venezuelae. It is 68 amino acid DNA-binding protein related to the MerR family of transcription factors. BldC deletion strains are bald because they initiate sporulation prematurely, omitting the formation of aerial hyphae altogether. This ChIP-Seq experiment was carried out to determine all the binding sites BldC binds to in the genome of Streptomyces venezuelae. Anti-BldC antibodies were used for ChIP-Seq of the wild type (WT) strain after 10 and 14 hours of growth in shaken cultures. A BldC deletion strain was made and anti-BldC antibodies were used for ChIP-Seq in the deletion strain after 14 hours of growth in shaken cultures. This was used as the negative control and enrichment in this ChIP-Seq was subtracted from the enrichment in the WT strain.
