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Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)

Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA(2)(Ile) to base pair specifically with the A of the AUA codon but not with the G of AUG. Recently, a Bacillus subtilis strain was isolated in which the essential...

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Autores principales: Köhrer, Caroline, Mandal, Debabrata, Gaston, Kirk W., Grosjean, Henri, Limbach, Patrick A., RajBhandary, Uttam L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2014
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3919564/
https://www.ncbi.nlm.nih.gov/pubmed/24194599
http://dx.doi.org/10.1093/nar/gkt1009
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author Köhrer, Caroline
Mandal, Debabrata
Gaston, Kirk W.
Grosjean, Henri
Limbach, Patrick A.
RajBhandary, Uttam L.
author_facet Köhrer, Caroline
Mandal, Debabrata
Gaston, Kirk W.
Grosjean, Henri
Limbach, Patrick A.
RajBhandary, Uttam L.
author_sort Köhrer, Caroline
collection PubMed
description Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA(2)(Ile) to base pair specifically with the A of the AUA codon but not with the G of AUG. Recently, a Bacillus subtilis strain was isolated in which the essential gene encoding tRNA(Ile)-lysidine synthetase was deleted for the first time. In such a strain, C34 at the wobble position of tRNA(2)(Ile) is expected to remain unmodified and cells depend on a mutant suppressor tRNA derived from tRNA(1)(Ile), in which G34 has been changed to U34. An important question, therefore, is how U34 base pairs with A without also base pairing with G. Here, we show (i) that unlike U34 at the wobble position of all B. subtilis tRNAs of known sequence, U34 in the mutant tRNA is not modified, and (ii) that the mutant tRNA binds strongly to the AUA codon on B. subtilis ribosomes but only weakly to AUG. These in vitro data explain why the suppressor strain displays only a low level of misreading AUG codons in vivo and, as shown here, grows at a rate comparable to that of the wild-type strain.
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spelling pubmed-39195642014-02-10 Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile) Köhrer, Caroline Mandal, Debabrata Gaston, Kirk W. Grosjean, Henri Limbach, Patrick A. RajBhandary, Uttam L. Nucleic Acids Res RNA Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA(2)(Ile) to base pair specifically with the A of the AUA codon but not with the G of AUG. Recently, a Bacillus subtilis strain was isolated in which the essential gene encoding tRNA(Ile)-lysidine synthetase was deleted for the first time. In such a strain, C34 at the wobble position of tRNA(2)(Ile) is expected to remain unmodified and cells depend on a mutant suppressor tRNA derived from tRNA(1)(Ile), in which G34 has been changed to U34. An important question, therefore, is how U34 base pairs with A without also base pairing with G. Here, we show (i) that unlike U34 at the wobble position of all B. subtilis tRNAs of known sequence, U34 in the mutant tRNA is not modified, and (ii) that the mutant tRNA binds strongly to the AUA codon on B. subtilis ribosomes but only weakly to AUG. These in vitro data explain why the suppressor strain displays only a low level of misreading AUG codons in vivo and, as shown here, grows at a rate comparable to that of the wild-type strain. Oxford University Press 2014-02 2013-11-03 /pmc/articles/PMC3919564/ /pubmed/24194599 http://dx.doi.org/10.1093/nar/gkt1009 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), 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
spellingShingle RNA
Köhrer, Caroline
Mandal, Debabrata
Gaston, Kirk W.
Grosjean, Henri
Limbach, Patrick A.
RajBhandary, Uttam L.
Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)
title Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)
title_full Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)
title_fullStr Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)
title_full_unstemmed Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)
title_short Life without tRNA(Ile)-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA(2)(Ile)
title_sort life without trna(ile)-lysidine synthetase: translation of the isoleucine codon aua in bacillus subtilis lacking the canonical trna(2)(ile)
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3919564/
https://www.ncbi.nlm.nih.gov/pubmed/24194599
http://dx.doi.org/10.1093/nar/gkt1009
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