Nitric Oxide Signaling for Actinorhodin Production in Streptomyces coelicolor A3(2) via the DevS/R Two-Component System

Abstract

Nitric oxide (NO) is an important signaling molecule in eukaryotic and prokaryotic cells. A previous study revealed an NOS-independent NO production metabolic cycle in which the three nitrogen oxides, nitrate (NO3-), nitrite (NO2-) and NO were generated in the actinobacterium Streptomyces coelicolor A3(2). NO was suggested to act as a signaling molecule, functioning as a hormone that regulates secondary metabolism. Here, we demonstrate the NO-mediated regulation of the production of the blue pigmented antibiotic, actinorhodin (ACT), via the heme-based DevS/R two-component system (TCS). Intracellular NO controls the stabilization or inactivation of DevS, depending on the NO concentration. An electrophoretic mobility shift assay and chromatin immunoprecipitation-quantitative polymerase chain reaction analysis revealed the direct binding between DevR and the promoter region of actII-ORF4, resulting in gene expression. Our results indicate that NO simultaneously regulates the DevS/R TCS, thereby strictly controlling the secondary metabolism of S. coelicolor A3(2). Importance Diverse organisms, such as mammals, plants, and bacteria, utilize NO via well known signal transduction mechanisms. Many useful secondary metabolites producer Streptomyces genus bacteria had been also suggested the metabolisms regulated by endogenously produced NO; however, the regulatory mechanisms remain to be elucidated. In this study, we demonstrated the molecular mechanism by which endogenously produced NO regulates antibiotic production via DevS/R TCS in S. coelicolor A3 (2). NO serves as both a stabilizer and a repressor in the regulation of antibiotic production. This report shows the mechanism by which Streptomyces utilizes endogenously produced NO to modulate their normal life cycle. Moreover, this study implies that studying NO signaling in actinobacteria can help in the development of both clinical strategies against pathogenic actinomycetes and the actinobacterial industries.

Differences at Species Level and in Repertoires of Secondary Metabolite Biosynthetic Gene Clusters among Streptomyces coelicolor A3(2) and Type Strains of S. coelicolor and Its Taxonomic Neighbors

“…Hatano et al [1] reclassified strain A3(2) from S. coelicolor to S. violaceoruber; however, this reclassification is not widely known. In addition to many recent reports [45][46][47][48], even GenBank still classifies strain A3(2) as S. coelicolor [49,50]. In this study, we showed that strain A3(2) can be reclassified to S. anthocyanicus based on DNA-DNA relatedness, because the whole-genome sequence of the type strain of S. violaceoruber has not yet been published.…”

Role of EmaSR in Ethanol Metabolism by Acinetobacter baumannii

“…Most TCS found in bacteria to date have the ability to regulate gene expression for antibiotic resistance [ 10 , 19 ], antibiotic production [ 20 ], virulence, biofilm formation and motility [ 21 ]. In addition, an increasing number of studies show that TCS are involved in metabolism; for example, the MxtR/ErdR TCS has been shown to be essential for acetate utilization in P. putida KT2440 [ 13 ].…”

Mutation of MtrA at the Predicted Phosphorylation Site Abrogates Its Role as a Global Regulator in Streptomyces venezuelae

“…Each sample was separated by SDS-PAGE using 12% polyacrylamide gels and transferred to Hybond ECL membranes (GE Amersham). The membranes were blocked with 5% fat-free milk at room temperature for 2 h, washed, and then incubated with anti-FLAG monoclonal antibody (1:3,000; Boster Biological Technology) at 4°C overnight ( 40 ). Then, the membranes were washed twice and incubated with the horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (H+L) secondary antibody (1:5,000; Boster Biological Technology) for 50 min at room temperature.…”