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The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis

Pathogenic bacteria, such as Yersinia pseudotuberculosis encounter reactive oxygen species (ROS) as one of the first lines of defense in the mammalian host. In return, the bacteria react by mounting an oxidative stress response. Previous global RNA structure probing studies provided evidence for tem...

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Autores principales: Scheller, Daniel, Becker, Franziska, Wimbert, Andrea, Meggers, Dominik, Pienkoß, Stephan, Twittenhoff, Christian, Knoke, Lisa R., Leichert, Lars I., Narberhaus, Franz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10358904/
https://www.ncbi.nlm.nih.gov/pubmed/37428814
http://dx.doi.org/10.1371/journal.pgen.1010669
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author Scheller, Daniel
Becker, Franziska
Wimbert, Andrea
Meggers, Dominik
Pienkoß, Stephan
Twittenhoff, Christian
Knoke, Lisa R.
Leichert, Lars I.
Narberhaus, Franz
author_facet Scheller, Daniel
Becker, Franziska
Wimbert, Andrea
Meggers, Dominik
Pienkoß, Stephan
Twittenhoff, Christian
Knoke, Lisa R.
Leichert, Lars I.
Narberhaus, Franz
author_sort Scheller, Daniel
collection PubMed
description Pathogenic bacteria, such as Yersinia pseudotuberculosis encounter reactive oxygen species (ROS) as one of the first lines of defense in the mammalian host. In return, the bacteria react by mounting an oxidative stress response. Previous global RNA structure probing studies provided evidence for temperature-modulated RNA structures in the 5’-untranslated region (5’-UTR) of various oxidative stress response transcripts, suggesting that opening of these RNA thermometer (RNAT) structures at host-body temperature relieves translational repression. Here, we systematically analyzed the transcriptional and translational regulation of ROS defense genes by RNA-sequencing, qRT-PCR, translational reporter gene fusions, enzymatic RNA structure probing and toeprinting assays. Transcription of four ROS defense genes was upregulated at 37°C. The trxA gene is transcribed into two mRNA isoforms, of which the most abundant short one contains a functional RNAT. Biochemical assays validated temperature-responsive RNAT-like structures in the 5’-UTRs of sodB, sodC and katA. However, they barely conferred translational repression in Y. pseudotuberculosis at 25°C suggesting partially open structures available to the ribosome in the living cell. Around the translation initiation region of katY we discovered a novel, highly efficient RNAT that was primarily responsible for massive induction of KatY at 37°C. By phenotypic characterization of catalase mutants and through fluorometric real-time measurements of the redox-sensitive roGFP2-Orp1 reporter in these strains, we revealed KatA as the primary H(2)O(2) scavenger. Consistent with the upregulation of katY, we observed an improved protection of Y. pseudotuberculosis at 37°C. Our findings suggest a multilayered regulation of the oxidative stress response in Yersinia and an important role of RNAT-controlled katY expression at host body temperature.
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spelling pubmed-103589042023-07-21 The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis Scheller, Daniel Becker, Franziska Wimbert, Andrea Meggers, Dominik Pienkoß, Stephan Twittenhoff, Christian Knoke, Lisa R. Leichert, Lars I. Narberhaus, Franz PLoS Genet Research Article Pathogenic bacteria, such as Yersinia pseudotuberculosis encounter reactive oxygen species (ROS) as one of the first lines of defense in the mammalian host. In return, the bacteria react by mounting an oxidative stress response. Previous global RNA structure probing studies provided evidence for temperature-modulated RNA structures in the 5’-untranslated region (5’-UTR) of various oxidative stress response transcripts, suggesting that opening of these RNA thermometer (RNAT) structures at host-body temperature relieves translational repression. Here, we systematically analyzed the transcriptional and translational regulation of ROS defense genes by RNA-sequencing, qRT-PCR, translational reporter gene fusions, enzymatic RNA structure probing and toeprinting assays. Transcription of four ROS defense genes was upregulated at 37°C. The trxA gene is transcribed into two mRNA isoforms, of which the most abundant short one contains a functional RNAT. Biochemical assays validated temperature-responsive RNAT-like structures in the 5’-UTRs of sodB, sodC and katA. However, they barely conferred translational repression in Y. pseudotuberculosis at 25°C suggesting partially open structures available to the ribosome in the living cell. Around the translation initiation region of katY we discovered a novel, highly efficient RNAT that was primarily responsible for massive induction of KatY at 37°C. By phenotypic characterization of catalase mutants and through fluorometric real-time measurements of the redox-sensitive roGFP2-Orp1 reporter in these strains, we revealed KatA as the primary H(2)O(2) scavenger. Consistent with the upregulation of katY, we observed an improved protection of Y. pseudotuberculosis at 37°C. Our findings suggest a multilayered regulation of the oxidative stress response in Yersinia and an important role of RNAT-controlled katY expression at host body temperature. Public Library of Science 2023-07-10 /pmc/articles/PMC10358904/ /pubmed/37428814 http://dx.doi.org/10.1371/journal.pgen.1010669 Text en © 2023 Scheller et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Scheller, Daniel
Becker, Franziska
Wimbert, Andrea
Meggers, Dominik
Pienkoß, Stephan
Twittenhoff, Christian
Knoke, Lisa R.
Leichert, Lars I.
Narberhaus, Franz
The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis
title The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis
title_full The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis
title_fullStr The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis
title_full_unstemmed The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis
title_short The oxidative stress response, in particular the katY gene, is temperature-regulated in Yersinia pseudotuberculosis
title_sort oxidative stress response, in particular the katy gene, is temperature-regulated in yersinia pseudotuberculosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10358904/
https://www.ncbi.nlm.nih.gov/pubmed/37428814
http://dx.doi.org/10.1371/journal.pgen.1010669
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