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Information on EC 2.1.1.256 - tRNA (guanine6-N2)-methyltransferase
for references in articles please use BRENDA:EC2.1.1.256
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EC Tree 2 Transferases
2.1 Transferring one-carbon groups
2.1.1 Methyltransferases
2.1.1.256 tRNA (guanine6-N2)-methyltransferase
IUBMB Comments
The enzyme specifically methylates guanine6 at N2 in tRNA.
The expected taxonomic range for this enzyme is: Eukaryota, Archaea, Bacteria
- 2.1.1.256
- nucleosides
-
n2-methylguanosine
-
pyrococcus
- thermophilus
-
thermus
- methyltransferases
-
thump
- furiosus
-
rossmann-fold
- guanosine
- trnaphe
-
trna-modifying
- mtases
Reaction Schemes
show+
guanine6 in tRNA
=
+
N2-methylguanine6 in tRNA
Synonyms
trm14, ttc1157, pf1002, m2g6 methyltransferase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
G6 MTase
m2G6 methyltransferase
m2G6 tRNA methylase
methyltransferase Trm14
Trm14
TrmN
tRNA (m2G6) methyltransferase
tRNA m2G6 MTase
tRNA:m2G6 methyltransferase
TTC1157
m2G6 methyltransferase
-
-
-
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methyltransferase Trm14
-
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + guanine6 in tRNA = S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
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-
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SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:tRNA (guanine6-N2)-methyltransferase
The enzyme specifically methylates guanine6 at N2 in tRNA.
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + guanine67 in tRNATrp
2 S-adenosyl-L-homocysteine + N2-dimethylguanine67 in tRNATrp
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine37 in tRNATrp
S-adenosyl-L-homocysteine + N2-methylguanine37 in tRNATrp
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine54 in tRNATrp
S-adenosyl-L-homocysteine + N2-methylguanine54 in tRNATrp
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
S-adenosyl-L-methionine + guanine6 in tRNA(Phe)
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA(Phe)
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
S-adenosyl-L-methionine + guanine67 in tRNATrp
S-adenosyl-L-homocysteine + N2-methylguanine67 in tRNATrp
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine8 in tRNATrp
S-adenosyl-L-homocysteine + N2-methylguanine8 in tRNATrp
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
Substrates: the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
Substrates: the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem. Contribution of the THUMP domain in tRNA recognition and catalysis, substrate binding and ligand-induced conformational changes in the RFM domain
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
Substrates: the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
Substrates: the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem. Contribution of the THUMP domain in tRNA recognition and catalysis, substrate binding and ligand-induced conformational changes in the RFM domain, overview
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA(Phe)
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA(Phe)
Substrates: production of a small amount of N2-dimethylguanine6 in tRNA(Phe)
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA(Phe)
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA(Phe)
Substrates: production of a small amount of N2-dimethylguanine6 in tRNA(Phe)
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA(Phe)
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA(Phe)
Substrates: production of a small amount of N2-dimethylguanine6 in tRNA(Phe)
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
Substrates: Trm14 generates m2G at position 6 in tRNACys
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
Substrates: wild-type or G6A/C67U tRNACys transcripts as substrates, Trm14 generates m2G at position 6 in tRNACys, i.e. tRNACys m2G6-forming activity
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
Substrates: Trm14 generates m2G at position 6 in tRNACys
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
Substrates: wild-type or G6A/C67U tRNACys transcripts as substrates, Trm14 generates m2G at position 6 in tRNACys, i.e. tRNACys m2G6-forming activity
Products: -
Products: -
?
additional information
?
-
Substrates: active site structure with bound ligands, S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, and sinefungin, overview
Products: -
Products: -
?
additional information
?
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Substrates: active site structure with bound ligands, S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, and sinefungin, overview
Products: -
Products: -
?
additional information
?
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Substrates: docking model of TrmN in complex with tRNAPhe of Thermus thermophilus
Products: -
Products: -
?
additional information
?
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Substrates: docking model of TrmN in complex with tRNAPhe of Thermus thermophilus
Products: -
Products: -
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
S-adenosyl-L-methionine + guanine67 in tRNATrp
S-adenosyl-L-homocysteine + N2-methylguanine67 in tRNATrp
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
Substrates: the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA
Substrates: the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
Substrates: Trm14 generates m2G at position 6 in tRNACys
Products: -
Products: -
?
S-adenosyl-L-methionine + guanine6 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNACys
Substrates: Trm14 generates m2G at position 6 in tRNACys
Products: -
Products: -
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
single-turnover, kinetic analysis, overview
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additional information
additional information
-
single-turnover, kinetic analysis, overview
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
additional information
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hyperthermophilic archaeon Thermococcus kodakarensis grows at 60-100°C
brenda
additional information
hyperthermophilic archaeon Thermococcus kodakarensis grows at 60-100°C
brenda
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
malfunction
physiological function
evolution
RNA MTases from the TrmN/Trm14 family are present in archaea, bacteria and eukaryota and all specifically modify tRNAPhe at guanosine 6 in the tRNA acceptor stem. RNA MTases can be classified into four superfamilies, overview
evolution
RNA MTases from the TrmN/Trm14 family are present in archaea, bacteria and eukaryota and all specifically modify tRNAPhe at guanosine 6 in the tRNA acceptor stem. RNA MTases can be classified into four superfamilies, overview
evolution
Trm14 is associated with cluster of orthologous groups, COG, 0116, and most closely resembles the m2G10 tRNA methylase Trm11. Phylogenetic analysis reveals a canonical archaeal/bacterial evolutionary separation with 20-30% sequence identities between the two branches, but it is likely that the detailed functions of COG 0116 enzymes differ between the archaeal and bacterial domains. Phylogenetic distribution of Trm14-like proteins, overview
evolution
a distinct tRNA 2'-O-methyltransferase, which methylates the 2'-OH of ribose at position 6 in tRNA and does not differentiate between adenine and cytosine, may exist in Thermococcus and Pyrococcus genera. In terms of the other enzymes responsible for the observed 2'-O-methylations, Cm56 is a product of Trm56 (EC 2.1.1.206)
evolution
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Trm14 is associated with cluster of orthologous groups, COG, 0116, and most closely resembles the m2G10 tRNA methylase Trm11. Phylogenetic analysis reveals a canonical archaeal/bacterial evolutionary separation with 20-30% sequence identities between the two branches, but it is likely that the detailed functions of COG 0116 enzymes differ between the archaeal and bacterial domains. Phylogenetic distribution of Trm14-like proteins, overview
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malfunction
inactivation of the trmN gene leads to a total absence of N2-methylguanine in tRNA but did not affect cell growth or the formation of other modified nucleosides in tRNA(Phe). Therefore, m2G6 does not appear to be involved in an essential function
malfunction
the lack of 2-methylguanosine (m2G) at position 67 in the trm11 trm14 double disruptant strain suggests that this methylation is mediated by m2G6 methyltransferase Trm14
malfunction
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inactivation of the trmN gene leads to a total absence of N2-methylguanine in tRNA but did not affect cell growth or the formation of other modified nucleosides in tRNA(Phe). Therefore, m2G6 does not appear to be involved in an essential function
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physiological function
tRNA m2G6 formation may play a role in stabilizing the structure of the RNA and function synergistically with these core-region modifications to stabilize the molecule against thermal stress. Also, Trm14-catalyzed m2G6 formation in tRNACys may play a role in modulating the aminoacylation efficiency of phosphoseryl-tRNA synthetase, i.e. SepRS. Control of SepRS (or CysRS) activity by SAM-dependent methylation of tRNACys might provide the methanogen cell with a regulatory mechanism by which protein synthesis rates can respond to environmental sulfur levels or to concentrations of sulfur metabolites
physiological function
the trm14 gene is responsible for the m2G67 modification. The modification site (G67) forms a Watson-Crick base pair with C6 in tRNA. Archaeal Trm14 methylates G6 in tRNA and contains a THUMP domain, which often recognizes the CCA terminus in tRNA. But it has not been confirmed that Thermococcus kodakarensis Trm14 methylates G6 in tRNA like Methanocaldococcus jannaschii Trm14. Furthermore, the presence of m2G6 modification in Thermococcus kodakarensis tRNAs is not confirmed. In total, trm14 is responsible for m2G at positions 8, 54, 37, and 67, respectively, as determined by analysis of tRNATrp from gene disruptant strains
physiological function
-
tRNA m2G6 formation may play a role in stabilizing the structure of the RNA and function synergistically with these core-region modifications to stabilize the molecule against thermal stress. Also, Trm14-catalyzed m2G6 formation in tRNACys may play a role in modulating the aminoacylation efficiency of phosphoseryl-tRNA synthetase, i.e. SepRS. Control of SepRS (or CysRS) activity by SAM-dependent methylation of tRNACys might provide the methanogen cell with a regulatory mechanism by which protein synthesis rates can respond to environmental sulfur levels or to concentrations of sulfur metabolites
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Show AA Sequence and Transmembrane Helices (193 entries)
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Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
additional information
additional information
the 381 amino acid Trm14 protein possesses a canonical RNA recognition THUMP domain at the N-terminus, followed by a gamma-class Rossmann fold amino-methyltransferase catalytic domain featuring the signature NPPY active site motif
additional information
-
the 381 amino acid Trm14 protein possesses a canonical RNA recognition THUMP domain at the N-terminus, followed by a gamma-class Rossmann fold amino-methyltransferase catalytic domain featuring the signature NPPY active site motif
additional information
-
the 381 amino acid Trm14 protein possesses a canonical RNA recognition THUMP domain at the N-terminus, followed by a gamma-class Rossmann fold amino-methyltransferase catalytic domain featuring the signature NPPY active site motif
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additional information
the enzyme consists of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase domain, with the main tRNA binding surface in a groove between the THUMP domain and the MTase domain, secondary sequence comparison, overview
additional information
-
the enzyme consists of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase domain, with the main tRNA binding surface in a groove between the THUMP domain and the MTase domain, secondary sequence comparison, overview
additional information
the enzyme consists of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase domain, with the main tRNA binding surface in a groove between the THUMP domain and the MTase domain, secondary sequence comparison, overview
additional information
-
the enzyme consists of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase domain, with the main tRNA binding surface in a groove between the THUMP domain and the MTase domain, secondary sequence comparison, overview
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant detagged enzyme in complex with S-adenosyl-L-methionine or reaction product S-adenosyl-homocysteine, and inhibitor sinefungin, hanging drop vapor diffusion, protein in 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 500 mM NaCl, 280 mM imidazole, and 1 mM DTT, mixing in a 1:1 ratio with crystallization solution containing 100 mM Tris-acetate, pH 8.0, 32% PEG 4000, and 15% glycerol, for the phasing, selenomethionine-derivatized PfTrm14 is crystallized in a crystallization solution containing 100 mM Tris-acetate, pH 8.0, 32% PEG 4000, 15% glycerol, after streak seeding from a native PfTrm14 crystal, crystal soaking in mother liquor containing 1 mM ligand, X-ray diffraction structure determination and analysis at 1.95-2.4 A resolution
purified recombinant His-tagged enzyme in complex with S-adenosyl-L-methionine or reaction product S-adenosyl-homocysteine, and inhibitor sinefungin, hanging drop vapor diffusion, protein in 50 mM Tris-HCl, pH 8.0, 250 mM NaCl, and 350 mM imidazole, mixing in a 1:1 ratio with crystallization solution 100 mM citrate/phosphate, pH 3.5, 15% PEG 6000, 200 mM NaCl, and 100 mM sodium citrate, crystal soaking in mother liquor containing 1 mM ligand, X-ray diffraction structure determination and analysis at 2.16 A resolution, solved in the apo state by molecular replacement to a resolution of 2.05 A
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
construction of a trm11 trm14 double disruptant strain that shows a lack of 2-methylguanosine (m2G) at position 67
additional information
construction of a trm11 trm14 double disruptant strain that shows a lack of 2-methylguanosine (m2G) at position 67
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli Rosetta (DE3)
expression of the N-terminally His-tagged emzyme in in Escherichia coli strain Rosetta (DE3)
expression of the N-terminally His-tagged enzyme in in Escherichia coli strain BL21 (DE3)
gene MJ0438 or trm14, DNA and amino acid sequence determination, analysis and comparisons, phylogenetic analysis
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Menezes, S.; Gaston, K.W.; Krivos, K.L.; Apolinario, E.E.; Reich, N.O.; Sowers, K.R.; Limbach, P.A.; Perona, J.J.
Formation of m2G6 in Methanocaldococcus jannaschii tRNA catalyzed by the novel methyltransferase Trm14
Nucleic Acids Res.
39
7641-7655
2011
Methanocaldococcus jannaschii (Q57880), Methanocaldococcus jannaschii, Methanocaldococcus jannaschii JAL-1 (Q57880)
brenda
Fislage, M.; Roovers, M.; Tuszynska, I.; Bujnicki, J.M.; Droogmans, L.; Versees, W.
Crystal structures of the tRNA:m2G6 methyltransferase Trm14/TrmN from two domains of life
Nucleic Acids Res.
40
5149-5161
2012
Pyrococcus furiosus (Q8U248), Pyrococcus furiosus, Thermus thermophilus (Q72IH5), Thermus thermophilus
brenda
Roovers, M.; Oudjama, Y.; Fislage, M.; Bujnicki, J.; Verses, W.; Droogmans, L.
The open reading frame TTC1157 of Thermus thermophilus HB27 encodes the methyltransferase forming N 2-methylguanosine at position 6 in tRNA
RNA
18
815-824
2012
Pyrococcus furiosus (Q8U248), Pyrococcus furiosus, Thermus thermophilus (Q72IH5), Thermus thermophilus, Thermus thermophilus DSM 7039 (Q72IH5)
brenda
Hirata, A.; Suzuki, T.; Nagano, T.; Fujii, D.; Okamoto, M.; Sora, M.; Lowe, T.M.; Kanai, T.; Atomi, H.; Suzuki, T.; Hori, H.
Distinct modified nucleosides in tRNATrp from thehyperthermophilic archaeon Thermococcus kodakarensis and requirement of tRNA m2G10/m22G10 methyltransferase (archaeal Trm11) for survival at high temperatures
J. Bacteriol.
201
e00448-19
2019
Thermococcus kodakarensis, Thermococcus kodakarensis (Q5JEM0)
brenda
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