Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527
Abstract Background We previously identified a regulator NsdAsr, which negatively regulated rimocidin biosynthesis in Streptomyces rimosus M527. However, the exact regulatory mechanism of NsdAsr on rimocidin production remains unknown. Results In this study, firstly, transcriptomic data demonstrated...
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BMC
2025-07-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-025-02784-z |
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| author | Yawen Xie Yujie Jiang Yongyong Zhang Andreas Bechthold Xiaoping Yu Zheng Ma |
| author_facet | Yawen Xie Yujie Jiang Yongyong Zhang Andreas Bechthold Xiaoping Yu Zheng Ma |
| author_sort | Yawen Xie |
| collection | DOAJ |
| description | Abstract Background We previously identified a regulator NsdAsr, which negatively regulated rimocidin biosynthesis in Streptomyces rimosus M527. However, the exact regulatory mechanism of NsdAsr on rimocidin production remains unknown. Results In this study, firstly, transcriptomic data demonstrated that the differentially expressed genes resulting from the over-expression of nsdA sr were primarily associated with several key metabolic pathways, including glycolysis, oxidative phosphorylation, and ribosome-related genes, all of which were downregulated. This directly impacted the concentrations of CoA and NADH, as confirmed by concentration measurement assays. Subsequently, the results of the ChIP-seq experiments revealed that NsdAsr directly binds to 49 target genes. Notably, these include RS18275 and RS18290 (both involved in fatty acid degradation) as well as rpoB (related to DNA transcription). The validity of the ChIP-seq assay for these three genes was further supported by in vitro electrophoretic mobility shift assays. Regarding RS18275 and RS18290, the results revealed that the binding of NsdAsr to these elements led to the downregulation of gene expression. This, in turn, resulted in a decrease in the levels of butyryl-CoA and malonyl-CoA, which are known precursors for rimocidin biosynthesis. Consequently, this negatively impacted on the biosynthesis of rimocidin. In the case of rpoB, the results indicated that NsdAsr binding led to a downregulation of overall protein levels. This was determined by enzymatic activity of report gene GUS and Western blot assay. Consequently, this resulted in a decrease in rimocidin yield. Conclusion This study reveals NsdAsr’s dual role in limiting rimocidin production by suppressing metabolic precursors and modulating protein expression. Integrated transcriptomic and ChIP-seq analyses provide critical insights into its regulatory mechanisms. |
| format | Article |
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| institution | DOAJ |
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| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
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| series | Microbial Cell Factories |
| spelling | doaj-art-3b82e8b4849341eeaf9f45e8e4b93ff62025-08-20T03:06:27ZengBMCMicrobial Cell Factories1475-28592025-07-0124111710.1186/s12934-025-02784-zStudy on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527Yawen Xie0Yujie Jiang1Yongyong Zhang2Andreas Bechthold3Xiaoping Yu4Zheng Ma5Key Laboratory of Microbiological Metrology, Measurement & Bio-Product Quality Security, State Administration for Market Regulation, College of Life Sciences, China Jiliang UniversityKey Laboratory of Microbiological Metrology, Measurement & Bio-Product Quality Security, State Administration for Market Regulation, College of Life Sciences, China Jiliang UniversityKey Laboratory of Microbiological Metrology, Measurement & Bio-Product Quality Security, State Administration for Market Regulation, College of Life Sciences, China Jiliang UniversityInstitute for Pharmaceutical Sciences, Pharmaceutical Biology and Biotechnology, University of FreiburgKey Laboratory of Microbiological Metrology, Measurement & Bio-Product Quality Security, State Administration for Market Regulation, College of Life Sciences, China Jiliang UniversityKey Laboratory of Microbiological Metrology, Measurement & Bio-Product Quality Security, State Administration for Market Regulation, College of Life Sciences, China Jiliang UniversityAbstract Background We previously identified a regulator NsdAsr, which negatively regulated rimocidin biosynthesis in Streptomyces rimosus M527. However, the exact regulatory mechanism of NsdAsr on rimocidin production remains unknown. Results In this study, firstly, transcriptomic data demonstrated that the differentially expressed genes resulting from the over-expression of nsdA sr were primarily associated with several key metabolic pathways, including glycolysis, oxidative phosphorylation, and ribosome-related genes, all of which were downregulated. This directly impacted the concentrations of CoA and NADH, as confirmed by concentration measurement assays. Subsequently, the results of the ChIP-seq experiments revealed that NsdAsr directly binds to 49 target genes. Notably, these include RS18275 and RS18290 (both involved in fatty acid degradation) as well as rpoB (related to DNA transcription). The validity of the ChIP-seq assay for these three genes was further supported by in vitro electrophoretic mobility shift assays. Regarding RS18275 and RS18290, the results revealed that the binding of NsdAsr to these elements led to the downregulation of gene expression. This, in turn, resulted in a decrease in the levels of butyryl-CoA and malonyl-CoA, which are known precursors for rimocidin biosynthesis. Consequently, this negatively impacted on the biosynthesis of rimocidin. In the case of rpoB, the results indicated that NsdAsr binding led to a downregulation of overall protein levels. This was determined by enzymatic activity of report gene GUS and Western blot assay. Consequently, this resulted in a decrease in rimocidin yield. Conclusion This study reveals NsdAsr’s dual role in limiting rimocidin production by suppressing metabolic precursors and modulating protein expression. Integrated transcriptomic and ChIP-seq analyses provide critical insights into its regulatory mechanisms.https://doi.org/10.1186/s12934-025-02784-zStreptomyces rimosusNsdAsrChIP-SeqPrecursor supplyGUS |
| spellingShingle | Yawen Xie Yujie Jiang Yongyong Zhang Andreas Bechthold Xiaoping Yu Zheng Ma Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527 Microbial Cell Factories Streptomyces rimosus NsdAsr ChIP-Seq Precursor supply GUS |
| title | Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527 |
| title_full | Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527 |
| title_fullStr | Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527 |
| title_full_unstemmed | Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527 |
| title_short | Study on the regulatory mechanism of NsdAsr on rimocidin biosynthesis in Streptomyces rimosus M527 |
| title_sort | study on the regulatory mechanism of nsdasr on rimocidin biosynthesis in streptomyces rimosus m527 |
| topic | Streptomyces rimosus NsdAsr ChIP-Seq Precursor supply GUS |
| url | https://doi.org/10.1186/s12934-025-02784-z |
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