A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei
Abstract Background Due to the important role of rifamycin in curing tuberculosis infection, the study on rifamycin has never been stopped. Although RifZ, which locates within the rifamycin biosynthetic cluster, has recently been characterized as a pathway-specific regulator for rifamycin biosynthes...
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| Language: | English |
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BMC
2018-01-01
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| Series: | Microbial Cell Factories |
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| Online Access: | http://link.springer.com/article/10.1186/s12934-018-0863-5 |
| _version_ | 1818500352565051392 |
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| author | Chao Lei Jingzhi Wang Yuanyuan Liu Xinqiang Liu Guoping Zhao Jin Wang |
| author_facet | Chao Lei Jingzhi Wang Yuanyuan Liu Xinqiang Liu Guoping Zhao Jin Wang |
| author_sort | Chao Lei |
| collection | DOAJ |
| description | Abstract Background Due to the important role of rifamycin in curing tuberculosis infection, the study on rifamycin has never been stopped. Although RifZ, which locates within the rifamycin biosynthetic cluster, has recently been characterized as a pathway-specific regulator for rifamycin biosynthesis, little is known about the regulation of rifamycin export. Results In this work, we proved that the expression of the rifamycin efflux pump (RifP) was regulated by RifQ, a TetR-family transcriptional regulator. Deletion of rifQ had little impact on bacterial growth, but resulted in improved rifamycin production, which was consistent with the reverse transcription PCR results that RifQ negatively regulated rifP’s transcription. With electrophoretic mobility shift assay and DNase I Footprinting assay, RifQ was found to directly bind to the promoter region of rifP, and a typical inverted repeat was identified within the RifQ-protected sequences. The transcription initiation site of rifP was further characterized and found to be upstream of the RifQ binding sites, well explaining the RifQ-mediated repression of rifP’s transcription in vivo. Moreover, rifamycin B (the end product of rifamycin biosynthesis) remarkably decreased the DNA binding affinity of RifQ, which led to derepression of rifamycin export, reducing the intracellular concentration of rifamycin B as well as its toxicity against the host. Conclusions Here, we proved that the export of rifamycin B was repressed by RifQ in Amycolatopsis mediterranei, and the RifQ-mediated repression could be specifically relieved by rifamycin B, the end product of rifamycin biosynthesis, based on which a feedback model was proposed for regulation of rifamycin export. With the findings here, one could improve the antibiotic yield by simply inactivating the negative regulator of the antibiotic transporter. |
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| language | English |
| last_indexed | 2024-12-10T20:41:32Z |
| publishDate | 2018-01-01 |
| publisher | BMC |
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| series | Microbial Cell Factories |
| spelling | doaj.art-efcab2feadaf4f37bfa3cc60cce5e0ec2022-12-22T01:34:22ZengBMCMicrobial Cell Factories1475-28592018-01-011711910.1186/s12934-018-0863-5A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterraneiChao Lei0Jingzhi Wang1Yuanyuan Liu2Xinqiang Liu3Guoping Zhao4Jin Wang5CAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesCAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesShanghai Tolo Biotechnology Company LimitedCAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesCAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesCAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesAbstract Background Due to the important role of rifamycin in curing tuberculosis infection, the study on rifamycin has never been stopped. Although RifZ, which locates within the rifamycin biosynthetic cluster, has recently been characterized as a pathway-specific regulator for rifamycin biosynthesis, little is known about the regulation of rifamycin export. Results In this work, we proved that the expression of the rifamycin efflux pump (RifP) was regulated by RifQ, a TetR-family transcriptional regulator. Deletion of rifQ had little impact on bacterial growth, but resulted in improved rifamycin production, which was consistent with the reverse transcription PCR results that RifQ negatively regulated rifP’s transcription. With electrophoretic mobility shift assay and DNase I Footprinting assay, RifQ was found to directly bind to the promoter region of rifP, and a typical inverted repeat was identified within the RifQ-protected sequences. The transcription initiation site of rifP was further characterized and found to be upstream of the RifQ binding sites, well explaining the RifQ-mediated repression of rifP’s transcription in vivo. Moreover, rifamycin B (the end product of rifamycin biosynthesis) remarkably decreased the DNA binding affinity of RifQ, which led to derepression of rifamycin export, reducing the intracellular concentration of rifamycin B as well as its toxicity against the host. Conclusions Here, we proved that the export of rifamycin B was repressed by RifQ in Amycolatopsis mediterranei, and the RifQ-mediated repression could be specifically relieved by rifamycin B, the end product of rifamycin biosynthesis, based on which a feedback model was proposed for regulation of rifamycin export. With the findings here, one could improve the antibiotic yield by simply inactivating the negative regulator of the antibiotic transporter.http://link.springer.com/article/10.1186/s12934-018-0863-5RifamycinAmycolatopsis mediterraneirifQExportFeedback regulation |
| spellingShingle | Chao Lei Jingzhi Wang Yuanyuan Liu Xinqiang Liu Guoping Zhao Jin Wang A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei Microbial Cell Factories Rifamycin Amycolatopsis mediterranei rifQ Export Feedback regulation |
| title | A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei |
| title_full | A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei |
| title_fullStr | A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei |
| title_full_unstemmed | A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei |
| title_short | A feedback regulatory model for RifQ-mediated repression of rifamycin export in Amycolatopsis mediterranei |
| title_sort | feedback regulatory model for rifq mediated repression of rifamycin export in amycolatopsis mediterranei |
| topic | Rifamycin Amycolatopsis mediterranei rifQ Export Feedback regulation |
| url | http://link.springer.com/article/10.1186/s12934-018-0863-5 |
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