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Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon
Archaeosine (G(+)), which is found only at position 15 in many archaeal tRNA, is formed by two steps, the replacement of the guanine base with preQ(0) by archaeosine tRNA-guanine transglycosylase (ArcTGT) and the subsequent modification of preQ(0) to G(+) by archaeosine synthase. However, tRNA(Leu)...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Oxford University Press
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770233/ https://www.ncbi.nlm.nih.gov/pubmed/26721388 http://dx.doi.org/10.1093/nar/gkv1522 |
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author | Kawamura, Takuya Hirata, Akira Ohno, Satoshi Nomura, Yuichiro Nagano, Tomoko Nameki, Nobukazu Yokogawa, Takashi Hori, Hiroyuki |
author_facet | Kawamura, Takuya Hirata, Akira Ohno, Satoshi Nomura, Yuichiro Nagano, Tomoko Nameki, Nobukazu Yokogawa, Takashi Hori, Hiroyuki |
author_sort | Kawamura, Takuya |
collection | PubMed |
description | Archaeosine (G(+)), which is found only at position 15 in many archaeal tRNA, is formed by two steps, the replacement of the guanine base with preQ(0) by archaeosine tRNA-guanine transglycosylase (ArcTGT) and the subsequent modification of preQ(0) to G(+) by archaeosine synthase. However, tRNA(Leu) from Thermoplasma acidophilum, a thermo-acidophilic archaeon, exceptionally has two G(+)13 and G(+)15 modifications. In this study, we focused on the biosynthesis mechanism of G(+)13 and G(+)15 modifications in this tRNA(Leu). Purified ArcTGT from Pyrococcus horikoshii, for which the tRNA recognition mechanism and structure were previously characterized, exchanged only the G15 base in a tRNA(Leu) transcript with (14)C-guanine. In contrast, T. acidophilum cell extract exchanged both G13 and G15 bases. Because T. acidophilum ArcTGT could not be expressed as a soluble protein in Escherichia coli, we employed an expression system using another thermophilic archaeon, Thermococcus kodakarensis. The arcTGT gene in T. kodakarensis was disrupted, complemented with the T. acidophilum arcTGT gene, and tRNA(Leu) variants were expressed. Mass spectrometry analysis of purified tRNA(Leu) variants revealed the modifications of G(+)13 and G(+)15 in the wild-type tRNA(Leu). Thus, T. acidophilum ArcTGT has a multisite specificity and is responsible for the formation of both G(+)13 and G(+)15 modifications. |
format | Online Article Text |
id | pubmed-4770233 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-47702332016-02-29 Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon Kawamura, Takuya Hirata, Akira Ohno, Satoshi Nomura, Yuichiro Nagano, Tomoko Nameki, Nobukazu Yokogawa, Takashi Hori, Hiroyuki Nucleic Acids Res RNA Archaeosine (G(+)), which is found only at position 15 in many archaeal tRNA, is formed by two steps, the replacement of the guanine base with preQ(0) by archaeosine tRNA-guanine transglycosylase (ArcTGT) and the subsequent modification of preQ(0) to G(+) by archaeosine synthase. However, tRNA(Leu) from Thermoplasma acidophilum, a thermo-acidophilic archaeon, exceptionally has two G(+)13 and G(+)15 modifications. In this study, we focused on the biosynthesis mechanism of G(+)13 and G(+)15 modifications in this tRNA(Leu). Purified ArcTGT from Pyrococcus horikoshii, for which the tRNA recognition mechanism and structure were previously characterized, exchanged only the G15 base in a tRNA(Leu) transcript with (14)C-guanine. In contrast, T. acidophilum cell extract exchanged both G13 and G15 bases. Because T. acidophilum ArcTGT could not be expressed as a soluble protein in Escherichia coli, we employed an expression system using another thermophilic archaeon, Thermococcus kodakarensis. The arcTGT gene in T. kodakarensis was disrupted, complemented with the T. acidophilum arcTGT gene, and tRNA(Leu) variants were expressed. Mass spectrometry analysis of purified tRNA(Leu) variants revealed the modifications of G(+)13 and G(+)15 in the wild-type tRNA(Leu). Thus, T. acidophilum ArcTGT has a multisite specificity and is responsible for the formation of both G(+)13 and G(+)15 modifications. Oxford University Press 2016-02-29 2015-12-31 /pmc/articles/PMC4770233/ /pubmed/26721388 http://dx.doi.org/10.1093/nar/gkv1522 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://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 |
spellingShingle | RNA Kawamura, Takuya Hirata, Akira Ohno, Satoshi Nomura, Yuichiro Nagano, Tomoko Nameki, Nobukazu Yokogawa, Takashi Hori, Hiroyuki Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon |
title | Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon |
title_full | Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon |
title_fullStr | Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon |
title_full_unstemmed | Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon |
title_short | Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon |
title_sort | multisite-specific archaeosine trna-guanine transglycosylase (arctgt) from thermoplasma acidophilum, a thermo-acidophilic archaeon |
topic | RNA |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770233/ https://www.ncbi.nlm.nih.gov/pubmed/26721388 http://dx.doi.org/10.1093/nar/gkv1522 |
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