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Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine
Dnmt2 methylates cytosine at position 38 of tRNA(Asp) in a variety of eukaryotic organisms. A correlation between the presence of the hypermodified nucleoside queuosine (Q) at position 34 of tRNA(Asp) and the Dnmt2 dependent C38 methylation was recently found in vivo for S. pombe and D. discoideum....
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5995894/ https://www.ncbi.nlm.nih.gov/pubmed/29892076 http://dx.doi.org/10.1038/s41598-018-27118-5 |
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author | Johannsson, Sven Neumann, Piotr Wulf, Alexander Welp, Luisa M. Gerber, Hans-Dieter Krull, Matthias Diederichsen, Ulf Urlaub, Henning Ficner, Ralf |
author_facet | Johannsson, Sven Neumann, Piotr Wulf, Alexander Welp, Luisa M. Gerber, Hans-Dieter Krull, Matthias Diederichsen, Ulf Urlaub, Henning Ficner, Ralf |
author_sort | Johannsson, Sven |
collection | PubMed |
description | Dnmt2 methylates cytosine at position 38 of tRNA(Asp) in a variety of eukaryotic organisms. A correlation between the presence of the hypermodified nucleoside queuosine (Q) at position 34 of tRNA(Asp) and the Dnmt2 dependent C38 methylation was recently found in vivo for S. pombe and D. discoideum. We demonstrate a direct effect of the Q-modification on the methyltransferase catalytic efficiency in vitro, as V(max)/K(0.5) of purified S. pombe Dnmt2 shows an increase for in vitro transcribed tRNA(Asp) containing Q34 to 6.27 ∗ 10(–3) s(−1) µM(−1) compared to 1.51 ∗ 10(–3) s(−1) µM(−1) for the unmodified substrate. Q34tRNA(Asp) exhibits an only slightly increased affinity for Dnmt2 in comparison to unmodified G34tRNA. In order to get insight into the structural basis for the Q-dependency, the crystal structure of S. pombe Dnmt2 was determined at 1.7 Å resolution. It closely resembles the known structures of human and E. histolytica Dnmt2, and contains the entire active site loop. The interaction with tRNA was analyzed by means of mass-spectrometry using UV cross-linked Dnmt2-tRNA complex. These cross-link data and computational docking of Dnmt2 and tRNA(Asp) reveal Q34 positioned adjacent to the S-adenosylmethionine occupying the active site, suggesting that the observed increase of Dnmt2 catalytic efficiency by queuine originates from optimal positioning of the substrate molecules and residues relevant for methyl transfer. |
format | Online Article Text |
id | pubmed-5995894 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-59958942018-06-21 Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine Johannsson, Sven Neumann, Piotr Wulf, Alexander Welp, Luisa M. Gerber, Hans-Dieter Krull, Matthias Diederichsen, Ulf Urlaub, Henning Ficner, Ralf Sci Rep Article Dnmt2 methylates cytosine at position 38 of tRNA(Asp) in a variety of eukaryotic organisms. A correlation between the presence of the hypermodified nucleoside queuosine (Q) at position 34 of tRNA(Asp) and the Dnmt2 dependent C38 methylation was recently found in vivo for S. pombe and D. discoideum. We demonstrate a direct effect of the Q-modification on the methyltransferase catalytic efficiency in vitro, as V(max)/K(0.5) of purified S. pombe Dnmt2 shows an increase for in vitro transcribed tRNA(Asp) containing Q34 to 6.27 ∗ 10(–3) s(−1) µM(−1) compared to 1.51 ∗ 10(–3) s(−1) µM(−1) for the unmodified substrate. Q34tRNA(Asp) exhibits an only slightly increased affinity for Dnmt2 in comparison to unmodified G34tRNA. In order to get insight into the structural basis for the Q-dependency, the crystal structure of S. pombe Dnmt2 was determined at 1.7 Å resolution. It closely resembles the known structures of human and E. histolytica Dnmt2, and contains the entire active site loop. The interaction with tRNA was analyzed by means of mass-spectrometry using UV cross-linked Dnmt2-tRNA complex. These cross-link data and computational docking of Dnmt2 and tRNA(Asp) reveal Q34 positioned adjacent to the S-adenosylmethionine occupying the active site, suggesting that the observed increase of Dnmt2 catalytic efficiency by queuine originates from optimal positioning of the substrate molecules and residues relevant for methyl transfer. Nature Publishing Group UK 2018-06-11 /pmc/articles/PMC5995894/ /pubmed/29892076 http://dx.doi.org/10.1038/s41598-018-27118-5 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Johannsson, Sven Neumann, Piotr Wulf, Alexander Welp, Luisa M. Gerber, Hans-Dieter Krull, Matthias Diederichsen, Ulf Urlaub, Henning Ficner, Ralf Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine |
title | Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine |
title_full | Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine |
title_fullStr | Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine |
title_full_unstemmed | Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine |
title_short | Structural insights into the stimulation of S. pombe Dnmt2 catalytic efficiency by the tRNA nucleoside queuosine |
title_sort | structural insights into the stimulation of s. pombe dnmt2 catalytic efficiency by the trna nucleoside queuosine |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5995894/ https://www.ncbi.nlm.nih.gov/pubmed/29892076 http://dx.doi.org/10.1038/s41598-018-27118-5 |
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