Project description:The aim of this experiment was to map the transcription start sites (TSSs) in the bottromycin biosynthetic gene cluster from Streptomyces scabies, qualitatively assess the expression levels of this cluster within the bacterium's transcriptome and evaluate whether deletion of a potential regulatory gene in the cluster, btmL affects gene cluster expression.
Project description:ForJ, ForF and ForZ are cluster situated regulators of the formicamycin biosynthetic gene cluster in Streptomyces forimcae. This ChIP Sequencing experiment was conducted to identify where in the genome these regulators bind in order to identify which transcripts they might regulate.
Project description:Here, we profiled the transcriptional capacity of a library of regulatory sequences mined from diverse Biosynthetic Gene Clusters in S. albidoflavus (S. albus J1074) to investigate BGC gene regulation.
Project description:The production of daptomycin (DAP) is precisely regulated by a complex regulatory network in Streptomyces roseosporus (S. roseosporus). Although the most biosynthetic pathway of DAP has been elucidated, the regulatory mechanism of its biosynthesis at transcriptional level is not fully understood. In the present study, a transcriptional regulator DhyR has been identified based on our previous quantification proteomics identification in S. roseosporus, and further the dhyR gene deletion mutant has been carried out to reveal DhyR function through transcriptome sequencing and loss-of function validation, which demonstrates DhyR positively regulates DAP biosynthesis in S. rosesporus. In-frame gene deletion of dhyR results in the significant downregulation of the transcription levels of all structural genes in DAP biosynthetic gene cluster and significantly decreases production of DAP. In contrast, overexpression of dhyR enhances transcription levels of DAP biosynthetic gene cluster and leads to a 23% increase of DAP yield. Moreover, the deletion of dhyR resultes in the significant down-regulation of transcription levels of another three regulatory genes, including atrA, depR1 and ssig-05090, which have been demonstrated to be closely associated with the DAP biosynthesis. Transcriptome analyses also support that DhyR modulates carbohydrate metabolism, amino acid metabolism and synthesis and transport pathways of siderophore in S. roseosporus. In summary, our findings indicate that DhyR functions as a pleiotropic regulator of primary and secondary metabolism to involve in DAP biosynthetic regulation of S. roseosporus.
