Information on EC 2.1.1.215 - tRNA (guanine26-N2/guanine27-N2)-dimethyltransferase

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The expected taxonomic range for this enzyme is: Bacteria, Archaea

EC NUMBER
COMMENTARY hide
2.1.1.215
-
RECOMMENDED NAME
GeneOntology No.
tRNA (guanine26-N2/guanine27-N2)-dimethyltransferase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4 S-adenosyl-L-methionine + guanine26/guanine27 in tRNA = 4 S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27 in tRNA
show the reaction diagram
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:tRNA (guanine26-N2/guanine27-N2)-dimethyltransferase
The enzyme from Aquifex aeolicus is similar to the TRM1 methyltransferases of archaea and eukarya (see EC 2.1.1.216, tRNA (guanine26-N2)-dimethyltransferase). However, it catalyses the double methylation of guanines at both positions 26 and 27 of tRNA.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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archaeal and eukaryotic tRNA (N2,N2-guanine)-dimethyltransferase, Trm1, produces N2,N2-dimethylguanine at position 26 in tRNA. In contrast, Trm1 from Aquifex aeolicus, a hyper-thermophilic eubacterium, modifies G27 as well as G26. The overall structure of Aquifex aeolicus Trm1 is similar to that of archaeal Trm1, although there is a zinc-cysteine cluster in the C-terminal domain of Aquifex aeolicus Trm1
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4 S-adenosyl-L-methionine + guanine26/guanine27 in tRNA
4 S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27 in tRNA
show the reaction diagram
4 S-adenosyl-L-methionine + guanine26/guanine27 in tRNACys
4 S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27 in tRNACys
show the reaction diagram
m2G26 formation is faster than the m2G27 formation and disruption of the G27-C43 base pair accelerates velocity of the G27 modification. A fraction of native tRNACys has a N2-dimethylguanine26/N2-dimethylguanine27 modification. Initially the N2-methylguanine26 modification occurs, and then the second methyl transfer reaction generates N2-dimethylguanine26. The third methyl transfer reaction modifies guanine 27 to N2-methylguanine27 followed by a fourth methylation of N2-methylguanine27 to N2-dimethylguanine27
-
-
?
4 S-adenosyl-L-methionine + guanine26/guanine27 in tRNAPhe
4 S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27 in tRNAPhe
show the reaction diagram
S-adenosyl-L-methionine + guanine26 in mutant tRNATyr
S-adenosyl-L-homocysteine + N2-methylguanine26 in mutant tRNATyr
show the reaction diagram
mutant tRNATyr in which the wilde-type adenine26/guanine27 sequence is substituted with guanine26/adenine27
-
-
?
S-adenosyl-L-methionine + guanine26/guanine27 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine26/guanine27 in tRNA
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + guanine26/guanine27 in tRNACys
S-adenosyl-L-homocysteine + N2-methylguanine26/guanine27 in tRNACys
show the reaction diagram
m2G26 formation is faster than the m2G27 formation and disruption of the G27-C43 base pair accelerates velocity of the G27 modification
-
-
?
S-adenosyl-L-methionine + guanine27 in tRNATyr
S-adenosyl-L-homocysteine + N2-methylguanine27 in tRNATyr
show the reaction diagram
wild-type tRNATyr contains the sequence adenine26/guanine27
-
-
?
S-adenosyl-L-methionine + N2-dimethylguanine26/guanine27 in tRNA
S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-methylguanine27
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + N2-dimethylguanine26/guanine27 in tRNACys
S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-methylguanine27 in tRNACys
show the reaction diagram
m2G26 formation is faster than the m2G27 formation and disruption of the G27-C43 base pair accelerates velocity of the G27 modification
-
-
?
S-adenosyl-L-methionine + N2-dimethylguanine26/N2-methylguanine27
S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + N2-dimethylguanine26/N2-methylguanine27 in tRNACys
S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27 in tRNACys
show the reaction diagram
m2G26 formation is faster than the m2G27 formation and disruption of the G27-C43 base pair accelerates velocity of the G27 modification
-
-
?
S-adenosyl-L-methionine + N2-methylguanine26 in mutant tRNATyr
S-adenosyl-L-homocysteine + N2-dimethylguanine26 in mutant tRNATyr
show the reaction diagram
mutant tRNATyr in which the wilde-type adenine26/guanine27 sequence is substituted with guanine26/adenine27
-
-
?
S-adenosyl-L-methionine + N2-methylguanine26/guanine27 in tRNA
S-adenosyl-L-homocysteine + N2-dimethylguanine26/guanine27 in tRNA
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + N2-methylguanine26/guanine27 in tRNACys
S-adenosyl-L-homocysteine + N2-dimethylguanine26/guanine27 in tRNACys
show the reaction diagram
m2G26 formation is faster than the m2G27 formation and disruption of the G27-C43 base pair accelerates velocity of the G27 modification
-
-
?
S-adenosyl-L-methionine + N2-methylguanine27 in tRNATyr
S-adenosyl-L-homocysteine + N2-dimethylguanine27 in tRNATyr
show the reaction diagram
wild-type tRNATyr contains the sequence adenine26/guanine27
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-
?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4 S-adenosyl-L-methionine + guanine26/guanine27 in tRNA
4 S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27 in tRNA
show the reaction diagram
S-adenosyl-L-methionine + guanine26/guanine27 in tRNA
S-adenosyl-L-homocysteine + N2-methylguanine26/guanine27 in tRNA
show the reaction diagram
O67010
-
-
-
?
S-adenosyl-L-methionine + N2-dimethylguanine26/guanine27 in tRNA
S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-methylguanine27
show the reaction diagram
O67010
-
-
-
?
S-adenosyl-L-methionine + N2-dimethylguanine26/N2-methylguanine27
S-adenosyl-L-homocysteine + N2-dimethylguanine26/N2-dimethylguanine27
show the reaction diagram
O67010
-
-
-
?
S-adenosyl-L-methionine + N2-methylguanine26/guanine27 in tRNA
S-adenosyl-L-homocysteine + N2-dimethylguanine26/guanine27 in tRNA
show the reaction diagram
O67010
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
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required
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
AdoEnYn
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a double-activated AdoMet analogue containing a pent-2-en-4-ynyl (EnYn) side chain instead of the methyl group. The double bond in beta-position to the sulfonium center counteracts unfavorable steric interactions within the SN2-like transition by conjugative stabilization and the terminal alkyne serves as reactive bio-orthogonal functionality. tRNAPhe methylation reaction to be inhibited by micromolar concentrations of AdoEnYn. EnYn-modified RNA structure determination by mass spectrometry
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00009
guanine26 in mutant tRNATyr
pH 7.5, 55°C, mutant tRNATyr in which the wilde-type adenine26/guanine27 sequence is substituted with guanine26/adenine27; pH 7.5, 55°C, mutant tRNATyr in which the wilde-type adenine26/guanine27 sequence is substituted with guanine26/adenine27, additional point mutation C43U
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0.00008 - 0.00017
guanine27 in tRNATyr
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0 - 0.33
S-adenosyl-L-methionine
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
AdoEnYn
Pyrococcus furiosus;
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pH and temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Aquifex aeolicus (strain VF5);
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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Trm1 from Aquifex aeolicus contains a zinc-cysteine cluster in the C-terminal domain. The N-terminal domain is a typical catalytic domain of S-adenosyl-L-methionine-dependent methyltransferases. Overall structure, structure comparisons, and structure-function analysis, overview
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged Trm1 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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expression in wheat germ
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expression of His6-tagged Trm1 in Escherichia coli strain BL21 (DE3)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D130A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D132A
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site-directed mutagenesis, inactive mutant
D84A
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site-directed mutagenesis, almost inactive mutant
E113A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E6A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F134A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F140A
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
F27A
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site-directed mutagenesis, almost inactive mutant
H110A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H219A
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
H274A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
I65A
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
I85A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
K170A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
K283A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
L60A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
N29A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R179A
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R192A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R31A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R36A
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site-directed mutagenesis, inactive mutant
R66A
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site-directed mutagenesis, almost inactive mutant