Crystal structures of the tRNA:m2G6 methyltransferase Trm14/TrmN from two domains of life.

Nucleic Acids Research
Marcus FislageWim Versées

Abstract

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 i...Continue Reading

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Citations

Aug 28, 2012·Nucleic Acids Research·Avinash S PunekarMaria Selmer
Aug 16, 2013·Nucleic Acids Research·Avinash S PunekarMaria Selmer
May 16, 2015·Nucleic Acids Research·Irina TuszynskaJanusz M Bujnicki
Jun 7, 2014·Frontiers in Genetics·Hiroyuki Hori
Jan 31, 2019·Nucleic Acids Research·Valérie de Crécy-LagardJanusz M Bujnicki
Mar 11, 2020·Microbial Biotechnology·Antoine DanchinConghui You
Oct 27, 2017·Journal of Biochemistry·Chie TomikawaHiroyuki Hori
Dec 18, 2016·Nucleic Acids Research·Gabrielle BourgeoisMarc Graille
Apr 4, 2020·Molecular Cell·Meredith CorleyGene W Yeo
Apr 22, 2021·Science China. Life Sciences·Jing LiRu-Juan Liu

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Methods Mentioned

BETA
X-ray
MTase
size exclusion chromatography
Band shift

Software Mentioned

PDBeFold
SHELXE
FILTREST3D
PhaserMR
wARP
Pymol
XDS
CCP4
PARSE
Molprobity

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