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Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice

Trehalose biosynthesis is considered an attractive target for the development of antimicrobials against fungal, helminthic and bacterial pathogens including Mycobacterium tuberculosis. The most common biosynthetic route involves trehalose-6-phosphate (T6P) synthase OtsA and T6P phosphatase OtsB that...

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Autores principales: Korte, Jan, Alber, Marina, Trujillo, Carolina M., Syson, Karl, Koliwer-Brandl, Hendrik, Deenen, René, Köhrer, Karl, DeJesus, Michael A., Hartman, Travis, Jacobs, William R., Bornemann, Stephen, Ioerger, Thomas R., Ehrt, Sabine, Kalscheuer, Rainer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5148154/
https://www.ncbi.nlm.nih.gov/pubmed/27936238
http://dx.doi.org/10.1371/journal.ppat.1006043
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author Korte, Jan
Alber, Marina
Trujillo, Carolina M.
Syson, Karl
Koliwer-Brandl, Hendrik
Deenen, René
Köhrer, Karl
DeJesus, Michael A.
Hartman, Travis
Jacobs, William R.
Bornemann, Stephen
Ioerger, Thomas R.
Ehrt, Sabine
Kalscheuer, Rainer
author_facet Korte, Jan
Alber, Marina
Trujillo, Carolina M.
Syson, Karl
Koliwer-Brandl, Hendrik
Deenen, René
Köhrer, Karl
DeJesus, Michael A.
Hartman, Travis
Jacobs, William R.
Bornemann, Stephen
Ioerger, Thomas R.
Ehrt, Sabine
Kalscheuer, Rainer
author_sort Korte, Jan
collection PubMed
description Trehalose biosynthesis is considered an attractive target for the development of antimicrobials against fungal, helminthic and bacterial pathogens including Mycobacterium tuberculosis. The most common biosynthetic route involves trehalose-6-phosphate (T6P) synthase OtsA and T6P phosphatase OtsB that generate trehalose from ADP/UDP-glucose and glucose-6-phosphate. In order to assess the drug target potential of T6P phosphatase, we generated a conditional mutant of M. tuberculosis allowing the regulated gene silencing of the T6P phosphatase gene otsB2. We found that otsB2 is essential for growth of M. tuberculosis in vitro as well as for the acute infection phase in mice following aerosol infection. By contrast, otsB2 is not essential for the chronic infection phase in mice, highlighting the substantial remodelling of trehalose metabolism during infection by M. tuberculosis. Blocking OtsB2 resulted in the accumulation of its substrate T6P, which appears to be toxic, leading to the self-poisoning of cells. Accordingly, blocking T6P production in a ΔotsA mutant abrogated otsB2 essentiality. T6P accumulation elicited a global upregulation of more than 800 genes, which might result from an increase in RNA stability implied by the enhanced neutralization of toxins exhibiting ribonuclease activity. Surprisingly, overlap with the stress response caused by the accumulation of another toxic sugar phosphate molecule, maltose-1-phosphate, was minimal. A genome-wide screen for synthetic lethal interactions with otsA identified numerous genes, revealing additional potential drug targets synergistic with OtsB2 suitable for combination therapies that would minimize the emergence of resistance to OtsB2 inhibitors.
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spelling pubmed-51481542016-12-28 Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice Korte, Jan Alber, Marina Trujillo, Carolina M. Syson, Karl Koliwer-Brandl, Hendrik Deenen, René Köhrer, Karl DeJesus, Michael A. Hartman, Travis Jacobs, William R. Bornemann, Stephen Ioerger, Thomas R. Ehrt, Sabine Kalscheuer, Rainer PLoS Pathog Research Article Trehalose biosynthesis is considered an attractive target for the development of antimicrobials against fungal, helminthic and bacterial pathogens including Mycobacterium tuberculosis. The most common biosynthetic route involves trehalose-6-phosphate (T6P) synthase OtsA and T6P phosphatase OtsB that generate trehalose from ADP/UDP-glucose and glucose-6-phosphate. In order to assess the drug target potential of T6P phosphatase, we generated a conditional mutant of M. tuberculosis allowing the regulated gene silencing of the T6P phosphatase gene otsB2. We found that otsB2 is essential for growth of M. tuberculosis in vitro as well as for the acute infection phase in mice following aerosol infection. By contrast, otsB2 is not essential for the chronic infection phase in mice, highlighting the substantial remodelling of trehalose metabolism during infection by M. tuberculosis. Blocking OtsB2 resulted in the accumulation of its substrate T6P, which appears to be toxic, leading to the self-poisoning of cells. Accordingly, blocking T6P production in a ΔotsA mutant abrogated otsB2 essentiality. T6P accumulation elicited a global upregulation of more than 800 genes, which might result from an increase in RNA stability implied by the enhanced neutralization of toxins exhibiting ribonuclease activity. Surprisingly, overlap with the stress response caused by the accumulation of another toxic sugar phosphate molecule, maltose-1-phosphate, was minimal. A genome-wide screen for synthetic lethal interactions with otsA identified numerous genes, revealing additional potential drug targets synergistic with OtsB2 suitable for combination therapies that would minimize the emergence of resistance to OtsB2 inhibitors. Public Library of Science 2016-12-09 /pmc/articles/PMC5148154/ /pubmed/27936238 http://dx.doi.org/10.1371/journal.ppat.1006043 Text en © 2016 Korte et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Korte, Jan
Alber, Marina
Trujillo, Carolina M.
Syson, Karl
Koliwer-Brandl, Hendrik
Deenen, René
Köhrer, Karl
DeJesus, Michael A.
Hartman, Travis
Jacobs, William R.
Bornemann, Stephen
Ioerger, Thomas R.
Ehrt, Sabine
Kalscheuer, Rainer
Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice
title Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice
title_full Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice
title_fullStr Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice
title_full_unstemmed Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice
title_short Trehalose-6-Phosphate-Mediated Toxicity Determines Essentiality of OtsB2 in Mycobacterium tuberculosis In Vitro and in Mice
title_sort trehalose-6-phosphate-mediated toxicity determines essentiality of otsb2 in mycobacterium tuberculosis in vitro and in mice
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5148154/
https://www.ncbi.nlm.nih.gov/pubmed/27936238
http://dx.doi.org/10.1371/journal.ppat.1006043
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