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, 55C, mutant tRNATyr in which the wilde-type adenine26/guanine27 sequence is substituted with guanine26/adenine27; pH 7.5, 55C, 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
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
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