Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2]
Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA[Ile over 2] 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 essent...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | en_US |
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Oxford University Press
2013
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| Online Access: | http://hdl.handle.net/1721.1/83245 https://orcid.org/0000-0002-4530-5647 |
| _version_ | 1811077396927873024 |
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| author | Koehrer, Caroline Mandal, Debabrata Gaston, Kirk W. Grosjean, Henri Limbach, Patrick A. Rajbhandary, Uttam L |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Koehrer, Caroline Mandal, Debabrata Gaston, Kirk W. Grosjean, Henri Limbach, Patrick A. Rajbhandary, Uttam L |
| author_sort | Koehrer, Caroline |
| collection | MIT |
| description | Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA[Ile over 2] 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[superscript Ile]-lysidine synthetase was deleted for the first time. In such a strain, C34 at the wobble position of tRNA[Ile over 2] is expected to remain unmodified and cells depend on a mutant suppressor tRNA derived from tRNA[Ile over 1], 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. |
| first_indexed | 2024-09-23T10:42:22Z |
| format | Article |
| id | mit-1721.1/83245 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:42:22Z |
| publishDate | 2013 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | mit-1721.1/832452022-09-30T22:22:08Z Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] Koehrer, Caroline Mandal, Debabrata Gaston, Kirk W. Grosjean, Henri Limbach, Patrick A. Rajbhandary, Uttam L Massachusetts Institute of Technology. Department of Biology Koehrer, Caroline Mandal, Debabrata Rajbhandary, Uttam L. Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA[Ile over 2] 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[superscript Ile]-lysidine synthetase was deleted for the first time. In such a strain, C34 at the wobble position of tRNA[Ile over 2] is expected to remain unmodified and cells depend on a mutant suppressor tRNA derived from tRNA[Ile over 1], 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. National Institutes of Health (U.S.) (GM17151) 2013-12-23T19:46:20Z 2013-12-23T19:46:20Z 2013-11 2013-10 Article http://purl.org/eprint/type/JournalArticle 0305-1048 1362-4962 http://hdl.handle.net/1721.1/83245 Kohrer, C., D. Mandal, K. W. Gaston, H. Grosjean, P. A. Limbach, and U. L. RajBhandary. “Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile.” Nucleic Acids Research (November 4, 2013). https://orcid.org/0000-0002-4530-5647 en_US http://dx.doi.org/10.1093/nar/gkt1009 Nucleic Acids Research http://creativecommons.org/licenses/by-nc/3.0/ application/pdf Oxford University Press Oxford University Press |
| spellingShingle | Koehrer, Caroline Mandal, Debabrata Gaston, Kirk W. Grosjean, Henri Limbach, Patrick A. Rajbhandary, Uttam L Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] |
| title | Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] |
| title_full | Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] |
| title_fullStr | Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] |
| title_full_unstemmed | Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] |
| title_short | Life without tRNA[superscript Ile]-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA[Ile over 2] |
| title_sort | life without trna superscript ile lysidine synthetase translation of the isoleucine codon aua in bacillus subtilis lacking the canonical trna ile over 2 |
| url | http://hdl.handle.net/1721.1/83245 https://orcid.org/0000-0002-4530-5647 |
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