Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>

ABSTRACT Lsr2 is a small nucleoid-associated protein found throughout the actinobacteria. Lsr2 functions similarly to the well-studied H-NS, in that it preferentially binds AT-rich sequences and represses gene expression. In Streptomyces venezuelae, Lsr2 represses the expression of many specialized...

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Autores principales:	Xiafei Zhang, Sara N. Andres, Marie A. Elliot
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	Streptomyces chloramphenicol antibiotic countersilencing nucleoid-associated protein Lsr2 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/eb90e39b313e49a499b6c2f469974153
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spelling	oai:doaj.org-article:eb90e39b313e49a499b6c2f4699741532021-11-10T18:37:50ZInterplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>10.1128/mBio.01077-212150-7511https://doaj.org/article/eb90e39b313e49a499b6c2f4699741532021-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01077-21https://doaj.org/toc/2150-7511ABSTRACT Lsr2 is a small nucleoid-associated protein found throughout the actinobacteria. Lsr2 functions similarly to the well-studied H-NS, in that it preferentially binds AT-rich sequences and represses gene expression. In Streptomyces venezuelae, Lsr2 represses the expression of many specialized metabolic clusters, including the chloramphenicol antibiotic biosynthetic gene cluster, and deleting lsr2 leads to significant upregulation of chloramphenicol cluster expression. We show here that Lsr2 likely exerts its repressive effects on the chloramphenicol cluster by polymerizing along the chromosome and by bridging sites within and adjacent to the chloramphenicol cluster. CmlR is a known activator of the chloramphenicol cluster, but expression of its associated gene is not upregulated in an lsr2 mutant strain. We demonstrate that CmlR is essential for chloramphenicol production, and further reveal that CmlR functions to “countersilence” Lsr2’s repressive effects by recruiting RNA polymerase and enhancing transcription, with RNA polymerase effectively clearing bound Lsr2 from the chloramphenicol cluster DNA. Our results provide insight into the interplay between opposing regulatory proteins that govern antibiotic production in S. venezuelae, which could be exploited to maximize the production of bioactive natural products in other systems. IMPORTANCE Specialized metabolic clusters in Streptomyces are the source of many clinically prescribed antibiotics. However, many clusters are not expressed in the laboratory due to repression by the nucleoid-associated protein Lsr2. Understanding how Lsr2 represses cluster expression, and how repression can be alleviated, is key to accessing the metabolic potential of these bacteria. Using the chloramphenicol biosynthetic cluster from Streptomyces venezuelae as a model, we explored the mechanistic basis underlying Lsr2-mediated repression, and activation by the pathway-specific regulator CmlR. Lsr2 polymerized along the chromosome and bridged binding sites located within and outside the cluster, promoting repression. Conversely, CmlR was essential for chloramphenicol production and further functioned to countersilence Lsr2 repression by recruiting RNA polymerase and promoting transcription, ultimately removing Lsr2 polymers from the chromosome. Manipulating the activity of both regulators led to a >130× increase in chloramphenicol levels, suggesting that combinatorial regulatory strategies can be powerful tools for maximizing natural product yields.Xiafei ZhangSara N. AndresMarie A. ElliotAmerican Society for MicrobiologyarticleStreptomyceschloramphenicolantibioticcountersilencingnucleoid-associated proteinLsr2MicrobiologyQR1-502ENmBio, Vol 12, Iss 4 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Streptomyces chloramphenicol antibiotic countersilencing nucleoid-associated protein Lsr2 Microbiology QR1-502
spellingShingle	Streptomyces chloramphenicol antibiotic countersilencing nucleoid-associated protein Lsr2 Microbiology QR1-502 Xiafei Zhang Sara N. Andres Marie A. Elliot Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>
description	ABSTRACT Lsr2 is a small nucleoid-associated protein found throughout the actinobacteria. Lsr2 functions similarly to the well-studied H-NS, in that it preferentially binds AT-rich sequences and represses gene expression. In Streptomyces venezuelae, Lsr2 represses the expression of many specialized metabolic clusters, including the chloramphenicol antibiotic biosynthetic gene cluster, and deleting lsr2 leads to significant upregulation of chloramphenicol cluster expression. We show here that Lsr2 likely exerts its repressive effects on the chloramphenicol cluster by polymerizing along the chromosome and by bridging sites within and adjacent to the chloramphenicol cluster. CmlR is a known activator of the chloramphenicol cluster, but expression of its associated gene is not upregulated in an lsr2 mutant strain. We demonstrate that CmlR is essential for chloramphenicol production, and further reveal that CmlR functions to “countersilence” Lsr2’s repressive effects by recruiting RNA polymerase and enhancing transcription, with RNA polymerase effectively clearing bound Lsr2 from the chloramphenicol cluster DNA. Our results provide insight into the interplay between opposing regulatory proteins that govern antibiotic production in S. venezuelae, which could be exploited to maximize the production of bioactive natural products in other systems. IMPORTANCE Specialized metabolic clusters in Streptomyces are the source of many clinically prescribed antibiotics. However, many clusters are not expressed in the laboratory due to repression by the nucleoid-associated protein Lsr2. Understanding how Lsr2 represses cluster expression, and how repression can be alleviated, is key to accessing the metabolic potential of these bacteria. Using the chloramphenicol biosynthetic cluster from Streptomyces venezuelae as a model, we explored the mechanistic basis underlying Lsr2-mediated repression, and activation by the pathway-specific regulator CmlR. Lsr2 polymerized along the chromosome and bridged binding sites located within and outside the cluster, promoting repression. Conversely, CmlR was essential for chloramphenicol production and further functioned to countersilence Lsr2 repression by recruiting RNA polymerase and promoting transcription, ultimately removing Lsr2 polymers from the chromosome. Manipulating the activity of both regulators led to a >130× increase in chloramphenicol levels, suggesting that combinatorial regulatory strategies can be powerful tools for maximizing natural product yields.
format	article
author	Xiafei Zhang Sara N. Andres Marie A. Elliot
author_facet	Xiafei Zhang Sara N. Andres Marie A. Elliot
author_sort	Xiafei Zhang
title	Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>
title_short	Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>
title_full	Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>
title_fullStr	Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>
title_full_unstemmed	Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>
title_sort	interplay between nucleoid-associated proteins and transcription factors in controlling specialized metabolism in <italic toggle="yes">streptomyces</italic>
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/eb90e39b313e49a499b6c2f469974153
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Interplay between Nucleoid-Associated Proteins and Transcription Factors in Controlling Specialized Metabolism in <italic toggle="yes">Streptomyces</italic>

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