A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>

ABSTRACT Many microorganisms produce secondary metabolites that have antibiotic activity. To avoid self-inhibition, the producing cells often encode cognate export and/or resistance mechanisms in the biosynthetic gene clusters for these molecules. Actinorhodin is a blue-pigmented antibiotic produced...

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Autores principales:	Ye Xu, Andrew Willems, Catherine Au-yeung, Kapil Tahlan, Justin R. Nodwell
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0de435cca3ca4bc693ec2a1ff6b2842b
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spelling	oai:doaj.org-article:0de435cca3ca4bc693ec2a1ff6b2842b2021-11-15T15:39:12ZA Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>10.1128/mBio.00191-122150-7511https://doaj.org/article/0de435cca3ca4bc693ec2a1ff6b2842b2012-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00191-12https://doaj.org/toc/2150-7511ABSTRACT Many microorganisms produce secondary metabolites that have antibiotic activity. To avoid self-inhibition, the producing cells often encode cognate export and/or resistance mechanisms in the biosynthetic gene clusters for these molecules. Actinorhodin is a blue-pigmented antibiotic produced by Streptomyces coelicolor. The actAB operon, carried in the actinorhodin biosynthetic gene cluster, encodes two putative export pumps and is regulated by the transcriptional repressor protein ActR. In this work, we show that normal actinorhodin yields require actAB expression. Consistent with previous in vitro work, we show that both actinorhodin and its 3-ring biosynthetic intermediates [e.g., (S)-DNPA] can relieve repression of actAB by ActR in vivo. Importantly, an ActR mutant that interacts productively with (S)-DNPA but not with actinorhodin responds to the actinorhodin biosynthetic pathway with the induction of actAB and normal yields of actinorhodin. This suggests that the intermediates are sufficient to trigger the export genes in actinorhodin-producing cells. We further show that actinorhodin-producing cells can induce actAB expression in nonproducing cells; however, in this case actinorhodin is the most important signal. Finally, while the “intermediate-only” ActR mutant permits sufficient actAB expression for normal actinorhodin yields, this expression is short-lived. Sustained culture-wide expression requires a subsequent actinorhodin-mediated signaling step, and the defect in this response causes widespread cell death. These results are consistent with a two-step model for actinorhodin export and resistance where intermediates trigger initial expression for export from producing cells and actinorhodin then triggers sustained export gene expression that confers culture-wide resistance. IMPORTANCE Understanding the links between antibiotic resistance and biosynthesis is important for our efforts to manipulate secondary metabolism. For example, many secondary metabolites are produced at low levels; our work suggests that manipulating export might be one way to enhance yields of these molecules. It also suggests that understanding resistance will be relevant to the generation of novel secondary metabolites through the creation of synthetic secondary metabolic gene clusters. Finally, these cognate resistance mechanisms are related to mechanisms that arise in pathogenic bacteria, and understanding them is relevant to our ability to control microbial infections clinically.Ye XuAndrew WillemsCatherine Au-yeungKapil TahlanJustin R. NodwellAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 5 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Ye Xu Andrew Willems Catherine Au-yeung Kapil Tahlan Justin R. Nodwell A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>
description	ABSTRACT Many microorganisms produce secondary metabolites that have antibiotic activity. To avoid self-inhibition, the producing cells often encode cognate export and/or resistance mechanisms in the biosynthetic gene clusters for these molecules. Actinorhodin is a blue-pigmented antibiotic produced by Streptomyces coelicolor. The actAB operon, carried in the actinorhodin biosynthetic gene cluster, encodes two putative export pumps and is regulated by the transcriptional repressor protein ActR. In this work, we show that normal actinorhodin yields require actAB expression. Consistent with previous in vitro work, we show that both actinorhodin and its 3-ring biosynthetic intermediates [e.g., (S)-DNPA] can relieve repression of actAB by ActR in vivo. Importantly, an ActR mutant that interacts productively with (S)-DNPA but not with actinorhodin responds to the actinorhodin biosynthetic pathway with the induction of actAB and normal yields of actinorhodin. This suggests that the intermediates are sufficient to trigger the export genes in actinorhodin-producing cells. We further show that actinorhodin-producing cells can induce actAB expression in nonproducing cells; however, in this case actinorhodin is the most important signal. Finally, while the “intermediate-only” ActR mutant permits sufficient actAB expression for normal actinorhodin yields, this expression is short-lived. Sustained culture-wide expression requires a subsequent actinorhodin-mediated signaling step, and the defect in this response causes widespread cell death. These results are consistent with a two-step model for actinorhodin export and resistance where intermediates trigger initial expression for export from producing cells and actinorhodin then triggers sustained export gene expression that confers culture-wide resistance. IMPORTANCE Understanding the links between antibiotic resistance and biosynthesis is important for our efforts to manipulate secondary metabolism. For example, many secondary metabolites are produced at low levels; our work suggests that manipulating export might be one way to enhance yields of these molecules. It also suggests that understanding resistance will be relevant to the generation of novel secondary metabolites through the creation of synthetic secondary metabolic gene clusters. Finally, these cognate resistance mechanisms are related to mechanisms that arise in pathogenic bacteria, and understanding them is relevant to our ability to control microbial infections clinically.
format	article
author	Ye Xu Andrew Willems Catherine Au-yeung Kapil Tahlan Justin R. Nodwell
author_facet	Ye Xu Andrew Willems Catherine Au-yeung Kapil Tahlan Justin R. Nodwell
author_sort	Ye Xu
title	A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>
title_short	A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>
title_full	A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>
title_fullStr	A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>
title_full_unstemmed	A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>
title_sort	two-step mechanism for the activation of actinorhodin export and resistance in <named-content content-type="genus-species">streptomyces coelicolor</named-content>
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/0de435cca3ca4bc693ec2a1ff6b2842b
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A Two-Step Mechanism for the Activation of Actinorhodin Export and Resistance in <named-content content-type="genus-species">Streptomyces coelicolor</named-content>

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