A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics

Expression of many bacterial genes is regulated by cis- and trans-acting elements in their 5’ upstream regions (URs). Cis-acting regulatory elements in URs include upstream ORFs (uORFs), short ORFs that sense translation stress that manifests as ribosomes stalling at specific codons within the uORF....

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Main Authors: Gabriele Baniulyte, Joseph T Wade
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2025-05-01
Series:eLife
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Online Access:https://elifesciences.org/articles/101217
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author Gabriele Baniulyte
Joseph T Wade
author_facet Gabriele Baniulyte
Joseph T Wade
author_sort Gabriele Baniulyte
collection DOAJ
description Expression of many bacterial genes is regulated by cis- and trans-acting elements in their 5’ upstream regions (URs). Cis-acting regulatory elements in URs include upstream ORFs (uORFs), short ORFs that sense translation stress that manifests as ribosomes stalling at specific codons within the uORF. Here, we show that the transcript encoding the Escherichia coli TopAI-YjhQ toxin–antitoxin system is regulated by a uORF that we name ‘toiL’. We propose that in the absence of translation stress, a secondary structure in the UR represses translation of the topAI transcript by occluding the ribosome-binding site. Translation repression of topAI leads to premature Rho-dependent transcription termination within the topAI ORF. At least five different classes of ribosome-targeting antibiotics relieve repression of topAI. Our data suggest that these antibiotics function by stalling ribosomes at different positions within toiL, thereby altering the RNA secondary structure around the topAI ribosome-binding site. Thus, toiL is a multipurpose uORF that can respond to a wide variety of translation stresses.
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spelling doaj-art-e5af5bb729814b0a97f525c3b8ae38dc2025-08-20T02:34:24ZengeLife Sciences Publications LtdeLife2050-084X2025-05-011310.7554/eLife.101217A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibioticsGabriele Baniulyte0https://orcid.org/0000-0003-0235-7938Joseph T Wade1https://orcid.org/0000-0002-9779-3160Department of Biomedical Sciences, School of Public Health, University at Albany, SUNY, Rensselaer, United StatesDepartment of Biomedical Sciences, School of Public Health, University at Albany, SUNY, Rensselaer, United States; Wadsworth Center, New York State Department of Health, Albany, United States; RNA Institute, University at Albany, SUNY, Albany, United StatesExpression of many bacterial genes is regulated by cis- and trans-acting elements in their 5’ upstream regions (URs). Cis-acting regulatory elements in URs include upstream ORFs (uORFs), short ORFs that sense translation stress that manifests as ribosomes stalling at specific codons within the uORF. Here, we show that the transcript encoding the Escherichia coli TopAI-YjhQ toxin–antitoxin system is regulated by a uORF that we name ‘toiL’. We propose that in the absence of translation stress, a secondary structure in the UR represses translation of the topAI transcript by occluding the ribosome-binding site. Translation repression of topAI leads to premature Rho-dependent transcription termination within the topAI ORF. At least five different classes of ribosome-targeting antibiotics relieve repression of topAI. Our data suggest that these antibiotics function by stalling ribosomes at different positions within toiL, thereby altering the RNA secondary structure around the topAI ribosome-binding site. Thus, toiL is a multipurpose uORF that can respond to a wide variety of translation stresses.https://elifesciences.org/articles/101217uORFtranslationRho-dependent transcription terminationribosome
spellingShingle Gabriele Baniulyte
Joseph T Wade
A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics
eLife
uORF
translation
Rho-dependent transcription termination
ribosome
title A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics
title_full A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics
title_fullStr A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics
title_full_unstemmed A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics
title_short A bacterial regulatory uORF senses multiple classes of ribosome-targeting antibiotics
title_sort bacterial regulatory uorf senses multiple classes of ribosome targeting antibiotics
topic uORF
translation
Rho-dependent transcription termination
ribosome
url https://elifesciences.org/articles/101217
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