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Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life

Methyltransferases (MTases) form a major class of tRNA-modifying enzymes needed for the proper functioning of tRNA. Recently, RNA MTases from the TrmN/Trm14 family that are present in Archaea, Bacteria and Eukaryota have been shown to specifically modify tRNA(Phe) at guanosine 6 in the tRNA acceptor...

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Autores principales: Fislage, Marcus, Roovers, Martine, Tuszynska, Irina, Bujnicki, Janusz M., Droogmans, Louis, Versées, Wim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3367198/
https://www.ncbi.nlm.nih.gov/pubmed/22362751
http://dx.doi.org/10.1093/nar/gks163
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author Fislage, Marcus
Roovers, Martine
Tuszynska, Irina
Bujnicki, Janusz M.
Droogmans, Louis
Versées, Wim
author_facet Fislage, Marcus
Roovers, Martine
Tuszynska, Irina
Bujnicki, Janusz M.
Droogmans, Louis
Versées, Wim
author_sort Fislage, Marcus
collection PubMed
description Methyltransferases (MTases) form a major class of tRNA-modifying enzymes needed for the proper functioning of tRNA. Recently, RNA MTases from the TrmN/Trm14 family that are present in Archaea, Bacteria and Eukaryota have been shown to specifically modify tRNA(Phe) at guanosine 6 in the tRNA acceptor stem. Here, we report the first X-ray crystal structures of the tRNA m(2)G6 (N(2)-methylguanosine) MTase (TTC)TrmN from Thermus thermophilus and its ortholog (Pf)Trm14 from Pyrococcus furiosus. Structures of (Pf)Trm14 were solved in complex with the methyl donor S-adenosyl-l-methionine (SAM or AdoMet), as well as the reaction product S-adenosyl-homocysteine (SAH or AdoHcy) and the inhibitor sinefungin. (TTC)TrmN and (Pf)Trm14 consist of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase (RFM) domain. These results represent the first crystallographic structure analysis of proteins containing both THUMP and RFM domain, and hence provide further insight in the contribution of the THUMP domain in tRNA recognition and catalysis. Electrostatics and conservation calculations suggest a main tRNA binding surface in a groove between the THUMP domain and the MTase domain. This is further supported by a docking model of TrmN in complex with tRNA(Phe) of T. thermophilus and via site-directed mutagenesis.
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spelling pubmed-33671982012-06-05 Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life Fislage, Marcus Roovers, Martine Tuszynska, Irina Bujnicki, Janusz M. Droogmans, Louis Versées, Wim Nucleic Acids Res Structural Biology Methyltransferases (MTases) form a major class of tRNA-modifying enzymes needed for the proper functioning of tRNA. Recently, RNA MTases from the TrmN/Trm14 family that are present in Archaea, Bacteria and Eukaryota have been shown to specifically modify tRNA(Phe) at guanosine 6 in the tRNA acceptor stem. Here, we report the first X-ray crystal structures of the tRNA m(2)G6 (N(2)-methylguanosine) MTase (TTC)TrmN from Thermus thermophilus and its ortholog (Pf)Trm14 from Pyrococcus furiosus. Structures of (Pf)Trm14 were solved in complex with the methyl donor S-adenosyl-l-methionine (SAM or AdoMet), as well as the reaction product S-adenosyl-homocysteine (SAH or AdoHcy) and the inhibitor sinefungin. (TTC)TrmN and (Pf)Trm14 consist of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase (RFM) domain. These results represent the first crystallographic structure analysis of proteins containing both THUMP and RFM domain, and hence provide further insight in the contribution of the THUMP domain in tRNA recognition and catalysis. Electrostatics and conservation calculations suggest a main tRNA binding surface in a groove between the THUMP domain and the MTase domain. This is further supported by a docking model of TrmN in complex with tRNA(Phe) of T. thermophilus and via site-directed mutagenesis. Oxford University Press 2012-06 2012-02-22 /pmc/articles/PMC3367198/ /pubmed/22362751 http://dx.doi.org/10.1093/nar/gks163 Text en © The Author(s) 2012. 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 unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Structural Biology
Fislage, Marcus
Roovers, Martine
Tuszynska, Irina
Bujnicki, Janusz M.
Droogmans, Louis
Versées, Wim
Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life
title Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life
title_full Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life
title_fullStr Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life
title_full_unstemmed Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life
title_short Crystal structures of the tRNA:m(2)G6 methyltransferase Trm14/TrmN from two domains of life
title_sort crystal structures of the trna:m(2)g6 methyltransferase trm14/trmn from two domains of life
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3367198/
https://www.ncbi.nlm.nih.gov/pubmed/22362751
http://dx.doi.org/10.1093/nar/gks163
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