GouR, a TetR Family Transcriptional Regulator, Coordinates the Biosynthesis and Export of Gougerotin in Streptomyces graminearus

Wei, Junhong; Tian, Yun; Niu, Guoqing; Tan, Huarong

doi:10.1128/aem.03003-13

Cited by 25 publications

(18 citation statements)

References 35 publications

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“…As a large and important family of one-component signal transduction systems, TFRs are widely associated with metabolism, antibiotic production, quorum sensing, and multidrug resistance (15,27). In the present study, we discovered that SLCG_2919 can regulate lincomycin biosynthesis by repressing the transcription of lincomycin biosynthetic structural genes, resistance genes, and a regulatory gene in the lin cluster, as well as by repressing its adjacent gene, SLCG_2920, which encodes an ATP/GTP binding protein.…”

Section: Discussionmentioning

confidence: 65%

TetR-Type Regulator SLCG_2919 Is a Negative Regulator of Lincomycin Biosynthesis in Streptomyces lincolnensis

et al. 2019

Appl Environ Microbiol

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Lincomycin A (Lin-A) is a widely used antibacterial antibiotic fermented by Streptomyces lincolnensis. However, the transcriptional regulatory mechanisms underlying lincomycin biosynthesis have seldom been investigated. Here, we first identified a TetR family transcriptional regulator (TFR), SLCG_2919, which negatively modulates lincomycin biosynthesis in S. lincolnensis LCGL. SLCG_2919 was found to specifically bind to promoter regions of the lincomycin biosynthetic gene cluster (lin cluster), including 25 structural genes, three resistance genes, and one regulatory gene, and to inhibit the transcription of these genes, demonstrating a directly regulatory role in lincomycin biosynthesis. Furthermore, we found that SLCG_2919 was not autoregulated, but directly repressed its adjacent gene, SLCG_2920, which encodes an ATP/GTP binding protein whose overexpression increased resistance against lincomycin and Lin-A yields in S. lincolnensis. The precise SLCG_2919 binding site within the promoter region of SLCG_2920 was determined by a DNase I footprinting assay and by electrophoretic mobility shift assays (EMSAs) based on base substitution mutagenesis, with the internal 10-nucleotide (nt) AT-rich sequence (AAATTATTTA) shown to be essential for SLCG_2919 binding. Our findings indicate that SLCG_2919 is a negative regulator for controlling lincomycin biosynthesis in S. lincolnensis. The present study improves our understanding of molecular regulation for lincomycin biosynthesis. IMPORTANCE TetR family transcriptional regulators (TFRs) are generally found to regulate diverse cellular processes in bacteria, especially antibiotic biosynthesis in Streptomyces species. However, knowledge of their function in lincomycin biosynthesis in S. lincolnensis remains unknown. The present study provides a new insight into the regulation of lincomycin biosynthesis through a TFR, SLCG_2919, that directly modulates lincomycin production and resistance. Intriguingly, SLCG_2919 and its adjoining gene, SLCG_2920, which encodes an ATP/GTP binding protein, were extensively distributed in diverse Streptomyces species. In addition, we revealed a new TFR binding motif, in which SLCG_2919 binds to the promoter region of SLCG_2920, dependent on the intervening AT-rich sequence rather than on the flanking inverted repeats found in the binding sites of other TFRs. These insights into transcriptional regulation of lincomycin biosynthesis by SLCG_2919 will be valuable in paving the way for genetic engineering of regulatory elements in Streptomyces species to improve antibiotic production.

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Section: Discussionmentioning

confidence: 65%

TetR-Type Regulator SLCG_2919 Is a Negative Regulator of Lincomycin Biosynthesis in Streptomyces lincolnensis

et al. 2019

Appl Environ Microbiol

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“…It is noteworthy that the PCC activity of the DBX(pccBX) strain was lower than that of the DF50 strain. The phenomenon of partial complementation of mutant phenotypes is widely found in different organisms (28). In the complemented DBX(pccBX) strain, the pccBX genes under the control of their native promoter were carried on a plasmid instead of being integrated into their original sites in the genome.…”

Section: Resultsmentioning

confidence: 99%

Propionyl Coenzyme A (Propionyl-CoA) Carboxylase in Haloferax mediterranei: Indispensability for Propionyl-CoA Assimilation and Impacts on Global Metabolism

Hou

Xiang

Han

2015

Appl Environ Microbiol

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Haloarchaea represent a distinct group of Archaea that typically inhabit hypersaline environments, in which nutrient supplies could vary considerably over time. Therefore, most of these extremophiles have developed the adaptation mechanism of depositing poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) intracellularly to store carbon and energy when carbon sources are oversupplied and utilizing PHBV in the absence of exogenous carbon sources (1, 2). During the process of PHBV biosynthesis and utilization, propionyl coenzyme A (propionyl-CoA) is an important intermediate metabolite. In addition, propionyl-CoA is also an essential intermediate of the methylaspartate cycle, a pathway for acetate assimilation in haloarchaea (3). In Bacteria and Eukarya, propionyl-CoA metabolism has been extensively studied, as excess propionyl-CoA inside the cell causes toxic effects (4, 5). For example, deficiencies in propionyl-CoA utilization affect polyketide synthesis, cell growth, and morphology of conidia of Aspergillus fumigatus (5) or lead to the serious disease propionic acidemia in humans (4). However, the enzymes for propionylCoA metabolism as well as the physiological roles of propionylCoA metabolism remain unclear for haloarchaea.Propionyl-CoA carboxylase (PCC) is the key enzyme for propionyl-CoA metabolism by catalyzing the carboxylation of propionyl-CoA to methylmalonyl-CoA. It is widely distributed in the three domains of life. PCC is typically composed of three functional components: the biotin carboxylase (BC), the biotin-carboxyl carrier protein (BCCP), and the carboxyltransferase (CT) (6). The crystal structure of a bacterial PCC holoenzyme has been reported, and a similar structure for human PCC has been obtained by using cryo-electron microscopy reconstruction (7). Recently, an acyl-CoA carboxylase with almost equal acetyl-CoA carboxylase (ACC) and PCC activities was characterized for the thermophilic archaeon Metallosphaera sedula (8). The ACC/PCC enzyme is responsible for two important carboxylation steps in the autotrophic carbon fixation cycle of the 3-hydroxypropionate/ 4-hydroxybutyrate cycle (9). Whether a similar ACC/PCC enzyme occurred in haloarchaea remained to be determined.Haloferax mediterranei is a metabolically versatile haloarchaeon and has been used as a model for studies of haloarchaeal metabolism, and especially PHBV biosynthesis, for decades (10). This strain can accumulate a large amount of PHBV with a high 3-hydroxyvalerate (3HV) content (ϳ10 mol%) from many unrelated carbon sources (11). In the absence of an exogenous carbon source, PHBV is degraded for nutritional purposes, and a large quantity of propionyl-CoA was able to be produced. Bioinformatic analysis revealed that potential genes encoding propionyl-

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“…Mfo1, a TetR family regulator, was found to negatively regulate the expression of mfs1 , which is consistent with previous observations that members of the TetR family of regulators can repress genes whose products are involved in multidrug resistance (Ramos et al ., ). In most cases, binding of TetR to its cognate DNA sequence could be dissociated by small‐molecule ligands, including antibiotic end products and their biosynthetic intermediates (Zhang et al ., ; Wei et al ., ). For instance, in biosynthesis of simocyclinone D8, the intermediate, simocyclinone C4, is effective in SimR dissociation in the regulation of the efflux pump gene simX (Le et al ., ).…”

Section: Discussionmentioning

confidence: 97%

Multiple transporters are involved in natamycin efflux in Streptomyces chattanoogensis L10

Wang

Shan

Han

et al. 2017

Molecular Microbiology

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Antibiotic-producing microorganisms have evolved several self-resistance mechanisms to prevent auto-toxicity. Overexpression of specific transporters to improve the efflux of toxic antibiotics has been found one of the most important and intrinsic resistance strategies used by many Streptomyces strains. In this work, two ATP-binding cassette (ABC) transporter-encoding genes located in the natamycin biosynthetic gene cluster, scnA and scnB, were identified as the primary exporter genes for natamycin efflux in Streptomyces chattanoogensis L10. Two other transporters located outside the cluster, a major facilitator superfamily transporter Mfs1 and an ABC transporter NepI/II were found to play a complementary role in natamycin efflux. ScnA/ScnB and Mfs1 also participate in exporting the immediate precursor of natamycin, 4,5-de-epoxynatamycin, which is more toxic to S. chattanoogensis L10 than natamycin. As the major complementary exporter for natamycin efflux, Mfs1 is up-regulated in response to intracellular accumulation of natamycin and 4,5-de-epoxynatamycin, suggesting a key role in the stress response for self-resistance. This article discusses a novel antibiotic-related efflux and response system in Streptomyces, as well as a self-resistance mechanism in antibiotic-producing strains.

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GouR, a TetR Family Transcriptional Regulator, Coordinates the Biosynthesis and Export of Gougerotin in Streptomyces graminearus

Cited by 25 publications

References 35 publications

TetR-Type Regulator SLCG_2919 Is a Negative Regulator of Lincomycin Biosynthesis in Streptomyces lincolnensis

TetR-Type Regulator SLCG_2919 Is a Negative Regulator of Lincomycin Biosynthesis in Streptomyces lincolnensis

Propionyl Coenzyme A (Propionyl-CoA) Carboxylase in Haloferax mediterranei: Indispensability for Propionyl-CoA Assimilation and Impacts on Global Metabolism

Multiple transporters are involved in natamycin efflux in Streptomyces chattanoogensis L10

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