A riboswitch separated from its ribosome-binding site still regulates translation
Riboswitches regulate downstream gene expression by binding cellular metabolites. Regulation of translation initiation by riboswitches is posited to occur by metabolite-mediated sequestration of the Shine-Dalgarno sequence (SDS), causing bypass by the ribosome. Recently, we solved a co-crystal struc...
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ftpubmed:oai:pubmedcentral.nih.gov:10018353 2023-05-15T14:13:32+02:00 A riboswitch separated from its ribosome-binding site still regulates translation Schroeder, Griffin M Akinyemi, Olayinka Malik, Jeffrey Focht, Caroline M Pritchett, Elizabeth M Baker, Cameron D McSally, James P Jenkins, Jermaine L Mathews, David H Wedekind, Joseph E 2023-02-10 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018353/ http://www.ncbi.nlm.nih.gov/pubmed/36762498 https://doi.org/10.1093/nar/gkad056 en eng Oxford University Press http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018353/ http://www.ncbi.nlm.nih.gov/pubmed/36762498 http://dx.doi.org/10.1093/nar/gkad056 © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Nucleic Acids Res Structural Biology Text 2023 ftpubmed https://doi.org/10.1093/nar/gkad056 2023-03-19T02:11:40Z Riboswitches regulate downstream gene expression by binding cellular metabolites. Regulation of translation initiation by riboswitches is posited to occur by metabolite-mediated sequestration of the Shine-Dalgarno sequence (SDS), causing bypass by the ribosome. Recently, we solved a co-crystal structure of a prequeuosine(1)-sensing riboswitch from Carnobacterium antarcticum that binds two metabolites in a single pocket. The structure revealed that the second nucleotide within the gene-regulatory SDS, G34, engages in a crystal contact, obscuring the molecular basis of gene regulation. Here, we report a co-crystal structure wherein C10 pairs with G34. However, molecular dynamics simulations reveal quick dissolution of the pair, which fails to reform. Functional and chemical probing assays inside live bacterial cells corroborate the dispensability of the C10–G34 pair in gene regulation, leading to the hypothesis that the compact pseudoknot fold is sufficient for translation attenuation. Remarkably, the C. antarcticum aptamer retained significant gene-regulatory activity when uncoupled from the SDS using unstructured spacers up to 10 nucleotides away from the riboswitch—akin to steric-blocking employed by sRNAs. Accordingly, our work reveals that the RNA fold regulates translation without SDS sequestration, expanding known riboswitch-mediated gene-regulatory mechanisms. The results infer that riboswitches exist wherein the SDS is not embedded inside a stable fold. Text Antarc* PubMed Central (PMC) Nucleic Acids Research 51 5 2464 2484 |
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Structural Biology |
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Structural Biology Schroeder, Griffin M Akinyemi, Olayinka Malik, Jeffrey Focht, Caroline M Pritchett, Elizabeth M Baker, Cameron D McSally, James P Jenkins, Jermaine L Mathews, David H Wedekind, Joseph E A riboswitch separated from its ribosome-binding site still regulates translation |
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Structural Biology |
description |
Riboswitches regulate downstream gene expression by binding cellular metabolites. Regulation of translation initiation by riboswitches is posited to occur by metabolite-mediated sequestration of the Shine-Dalgarno sequence (SDS), causing bypass by the ribosome. Recently, we solved a co-crystal structure of a prequeuosine(1)-sensing riboswitch from Carnobacterium antarcticum that binds two metabolites in a single pocket. The structure revealed that the second nucleotide within the gene-regulatory SDS, G34, engages in a crystal contact, obscuring the molecular basis of gene regulation. Here, we report a co-crystal structure wherein C10 pairs with G34. However, molecular dynamics simulations reveal quick dissolution of the pair, which fails to reform. Functional and chemical probing assays inside live bacterial cells corroborate the dispensability of the C10–G34 pair in gene regulation, leading to the hypothesis that the compact pseudoknot fold is sufficient for translation attenuation. Remarkably, the C. antarcticum aptamer retained significant gene-regulatory activity when uncoupled from the SDS using unstructured spacers up to 10 nucleotides away from the riboswitch—akin to steric-blocking employed by sRNAs. Accordingly, our work reveals that the RNA fold regulates translation without SDS sequestration, expanding known riboswitch-mediated gene-regulatory mechanisms. The results infer that riboswitches exist wherein the SDS is not embedded inside a stable fold. |
format |
Text |
author |
Schroeder, Griffin M Akinyemi, Olayinka Malik, Jeffrey Focht, Caroline M Pritchett, Elizabeth M Baker, Cameron D McSally, James P Jenkins, Jermaine L Mathews, David H Wedekind, Joseph E |
author_facet |
Schroeder, Griffin M Akinyemi, Olayinka Malik, Jeffrey Focht, Caroline M Pritchett, Elizabeth M Baker, Cameron D McSally, James P Jenkins, Jermaine L Mathews, David H Wedekind, Joseph E |
author_sort |
Schroeder, Griffin M |
title |
A riboswitch separated from its ribosome-binding site still regulates translation |
title_short |
A riboswitch separated from its ribosome-binding site still regulates translation |
title_full |
A riboswitch separated from its ribosome-binding site still regulates translation |
title_fullStr |
A riboswitch separated from its ribosome-binding site still regulates translation |
title_full_unstemmed |
A riboswitch separated from its ribosome-binding site still regulates translation |
title_sort |
riboswitch separated from its ribosome-binding site still regulates translation |
publisher |
Oxford University Press |
publishDate |
2023 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018353/ http://www.ncbi.nlm.nih.gov/pubmed/36762498 https://doi.org/10.1093/nar/gkad056 |
genre |
Antarc* |
genre_facet |
Antarc* |
op_source |
Nucleic Acids Res |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018353/ http://www.ncbi.nlm.nih.gov/pubmed/36762498 http://dx.doi.org/10.1093/nar/gkad056 |
op_rights |
© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
op_doi |
https://doi.org/10.1093/nar/gkad056 |
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Nucleic Acids Research |
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51 |
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5 |
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2464 |
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2484 |
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1766285994362929152 |