Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S
Abstract Background The anthraquinone-fused 10-membered enediynes (AFEs), represented by tiancimycins (TNMs), possess a unique structural feature and promising potentials as payloads of antitumor antibody–drug conjugates. Despite many efforts, the insufficient yields remain a practical challenge for...
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| Format: | Article |
| Language: | English |
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BMC
2022-09-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-022-01916-z |
| _version_ | 1811273634558246912 |
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| author | Manxiang Zhu Fan Zhang Ting Gan Jing Lin Yanwen Duan Xiangcheng Zhu |
| author_facet | Manxiang Zhu Fan Zhang Ting Gan Jing Lin Yanwen Duan Xiangcheng Zhu |
| author_sort | Manxiang Zhu |
| collection | DOAJ |
| description | Abstract Background The anthraquinone-fused 10-membered enediynes (AFEs), represented by tiancimycins (TNMs), possess a unique structural feature and promising potentials as payloads of antitumor antibody–drug conjugates. Despite many efforts, the insufficient yields remain a practical challenge for development of AFEs. Recent studies have suggested a unified basic biosynthetic route for AFEs, those core genes involved in the formation of essential common AFE intermediates, together with multiple regulatory genes, are highly conserved among the reported biosynthetic gene clusters (BGCs) of AFEs. The extreme cytotoxicities of AFEs have compelled hosts to evolve strict regulations to control their productions, but the exact roles of related regulatory genes are still uncertain. Results In this study, the genetic validations of five putative regulatory genes present in the BGC of TNMs revealed that only three (tnmR1, tnmR3 and tnmR7) of them were involved in the regulation of TNMs biosynthesis. The bioinformatic analysis also revealed that they represented three major but distinct groups of regulatory genes conserved in all BGCs of AFEs. Further transcriptional analyses suggested that TnmR7 could promote the expressions of core enzymes TnmD/G and TnmN/O/P, while TnmR3 may act as a sensor kinase to work with TnmR1 and form a higher class unconventional orphan two-component regulatory system, which dynamically represses the expressions of TnmR7, core enzymes TnmD/G/J/K1/K2 and auxiliary proteins TnmT2/S2/T1/S1. Therefore, the biosynthesis of TNMs was stringently restricted by this cascade regulatory network at early stage to ensure the normal cell growth, and then partially released at the stationary phase for product accumulation. Conclusion The pathway-specific cascade regulatory network consisting with TnmR3/R1 and TnmR7 was deciphered to orchestrate the production of TNMs. And it could be speculated as a common regulatory mechanism for productions of AFEs, which shall provide us new insights in future titer improvement of AFEs and potential dynamic regulatory applications in synthetic biology. |
| first_indexed | 2024-04-12T23:02:53Z |
| format | Article |
| id | doaj.art-db7d1f6315854f2f9f6d70f86b72a230 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-12T23:02:53Z |
| publishDate | 2022-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-db7d1f6315854f2f9f6d70f86b72a2302022-12-22T03:13:01ZengBMCMicrobial Cell Factories1475-28592022-09-0121111110.1186/s12934-022-01916-zDeciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-SManxiang Zhu0Fan Zhang1Ting Gan2Jing Lin3Yanwen Duan4Xiangcheng Zhu5Xiangya International Academy of Translational Medicine, Central South UniversityXiangya International Academy of Translational Medicine, Central South UniversityXiangya International Academy of Translational Medicine, Central South UniversityXiangya International Academy of Translational Medicine, Central South UniversityXiangya International Academy of Translational Medicine, Central South UniversityXiangya International Academy of Translational Medicine, Central South UniversityAbstract Background The anthraquinone-fused 10-membered enediynes (AFEs), represented by tiancimycins (TNMs), possess a unique structural feature and promising potentials as payloads of antitumor antibody–drug conjugates. Despite many efforts, the insufficient yields remain a practical challenge for development of AFEs. Recent studies have suggested a unified basic biosynthetic route for AFEs, those core genes involved in the formation of essential common AFE intermediates, together with multiple regulatory genes, are highly conserved among the reported biosynthetic gene clusters (BGCs) of AFEs. The extreme cytotoxicities of AFEs have compelled hosts to evolve strict regulations to control their productions, but the exact roles of related regulatory genes are still uncertain. Results In this study, the genetic validations of five putative regulatory genes present in the BGC of TNMs revealed that only three (tnmR1, tnmR3 and tnmR7) of them were involved in the regulation of TNMs biosynthesis. The bioinformatic analysis also revealed that they represented three major but distinct groups of regulatory genes conserved in all BGCs of AFEs. Further transcriptional analyses suggested that TnmR7 could promote the expressions of core enzymes TnmD/G and TnmN/O/P, while TnmR3 may act as a sensor kinase to work with TnmR1 and form a higher class unconventional orphan two-component regulatory system, which dynamically represses the expressions of TnmR7, core enzymes TnmD/G/J/K1/K2 and auxiliary proteins TnmT2/S2/T1/S1. Therefore, the biosynthesis of TNMs was stringently restricted by this cascade regulatory network at early stage to ensure the normal cell growth, and then partially released at the stationary phase for product accumulation. Conclusion The pathway-specific cascade regulatory network consisting with TnmR3/R1 and TnmR7 was deciphered to orchestrate the production of TNMs. And it could be speculated as a common regulatory mechanism for productions of AFEs, which shall provide us new insights in future titer improvement of AFEs and potential dynamic regulatory applications in synthetic biology.https://doi.org/10.1186/s12934-022-01916-zAnthraquinone-fused enediynesTiancimycinsOrphan two-component regulatory systemPathway-specific cascade regulatory networkEMSAqRT-PCR |
| spellingShingle | Manxiang Zhu Fan Zhang Ting Gan Jing Lin Yanwen Duan Xiangcheng Zhu Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S Microbial Cell Factories Anthraquinone-fused enediynes Tiancimycins Orphan two-component regulatory system Pathway-specific cascade regulatory network EMSA qRT-PCR |
| title | Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S |
| title_full | Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S |
| title_fullStr | Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S |
| title_full_unstemmed | Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S |
| title_short | Deciphering the pathway-specific regulatory network for production of ten-membered enediyne Tiancimycins in Streptomyces sp. CB03234-S |
| title_sort | deciphering the pathway specific regulatory network for production of ten membered enediyne tiancimycins in streptomyces sp cb03234 s |
| topic | Anthraquinone-fused enediynes Tiancimycins Orphan two-component regulatory system Pathway-specific cascade regulatory network EMSA qRT-PCR |
| url | https://doi.org/10.1186/s12934-022-01916-z |
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