Information on EC 2.1.1.57 - methyltransferase cap1

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.57
-
RECOMMENDED NAME
GeneOntology No.
methyltransferase cap1
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + a 5'-(N7-methyl 5'-triphosphoguanosine)-(purine-ribonucleotide)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N7-methyl 5'-triphosphoguanosine)-(2'-O-methyl-purine-ribonucleotide)-[mRNA]
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
methyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
mRNA capping II
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:5-(N7-methyl 5-triphosphoguanosine)-(purine-ribonucleotide)-[mRNA] 2-O-methyltransferase
This enzyme catalyses the methylation of the ribose on the first transcribed nucleotide of mRNA or snRNA molecules, which may be either guanosine or adenosine. This methylation event is known as cap1, and occurrs in all mRNAs and snRNAs of higher eukaryotes, including insects, vertebrates and their viruses. The human enzyme can also methylate mRNA molecules that lack methylation on the capping 5'-triphosphoguanosine [6].
CAS REGISTRY NUMBER
COMMENTARY hide
61970-02-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
viruses lacking 2'-O methyltransferase activity exhibit attenuation in primary macrophages
physiological function
-
the enzyme is responsible for cap1 formation in vivo
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + 7Me-GpppA-C5
S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
show the reaction diagram
S-adenosyl-L-methionine + 7Me-GpppA-Cn
S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-Cn
show the reaction diagram
-
exclusive methylation at the 2'O position
-
-
?
S-adenosyl-L-methionine + 7Me-GpppG
S-adenosyl-L-homocysteine + 7Me-Gppp-2'OMe-G
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + ApppGR-RNA
?
show the reaction diagram
S-adenosyl-L-methionine + GpppA-Cn
S-adenosyl-L-homocysteine + GpppA-2'OMe-Cn
show the reaction diagram
-
exclusive methylation at the 2'O position
-
-
?
S-adenosyl-L-methionine + GpppACCCCC
S-adenosyl-L-homocysteine + GpppAmCCCCC
show the reaction diagram
-
-
?
S-adenosyl-L-methionine + m7G(5')pppA-RNA
S-adenosyl-L-homocysteine + m7G(5')pppAm-RNA
show the reaction diagram
S-adenosyl-L-methionine + m7G(5')pppA-RNA
S-adenosyl-L-homocysteine + m7G(5')pppRA-RNA
show the reaction diagram
S-adenosyl-L-methionine + m7G(5')pppG-RNA
S-adenosyl-L-homocysteine + m7G(5')pppGm-RNA
show the reaction diagram
S-adenosyl-L-methionine + m7G(5')pppN-RNA
S-adenosyl-L-homocysteine + m7G(5')pppNm-RNA
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + m7G(5')pppNmpN-RNA
S-adenosyl-L-homocysteine + m7G(5')pppNmpNm-RNA
show the reaction diagram
-
N: purin and pyrimidine nucleotides are methylated, enzyme: cap II-methyltransferase
i.e. cap II
?
S-adenosyl-L-methionine + m7G(5')pppNpN-RNA
S-adenosyl-L-homocysteine + m7G(5')pppNmpN-RNA
show the reaction diagram
-
i.e. cap O, N: purine and pyrimidine nucleotides are methylated, enzyme: cap I-methyltransferase, substrates are RNAs with a capped terminus with at least 2 additional nucleotides, G(5')pppNpNp
i.e. cap I
?
S-adenosyl-L-methionine + m7G(5')pppR-RNA
S-adenosyl-L-homocysteine + m7G(5')pppRm-RNA
show the reaction diagram
S-adenosyl-L-methionine + m7G-(5')pppGpApApA
?
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + m7GpppACCCCC
S-adenosyl-L-homocysteine + m7GpppAmCCCCC
show the reaction diagram
-
-
?
S-adenosyl-L-methionine + m7GpppApGp
S-adenosyl-L-homocysteine + m7GpppAmpGp
show the reaction diagram
-
i.e. capped dinucleotide, cap I-methyltransferase
-
?
S-adenosyl-L-methionine + m7GpppApGpUp
S-adenosyl-L-homocysteine + m7GpppAmpGpUp
show the reaction diagram
-
i.e. capped trinucleotide, cap I-methyltransferase
-
?
S-adenosyl-L-methionine + m7GpppG
S-adenosyl-L-homocysteine + m7GpppGm
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + m7GpppGp
S-adenosyl-L-homocysteine + m7GpppGmp
show the reaction diagram
-
substrate of minimal chain length for the cap-specific enzyme, m7GpppG is no substrate
-
-
?
S-adenosyl-L-methionine + m7GpppGpN
S-adenosyl-L-homocysteine + m7GpppGmpN
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpNp
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpUbiotin-p
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + mononucleotide
?
show the reaction diagram
-
such as m7GpppA or GpppA, poor substrates for cap I-methyltransferase
-
-
?
additional information
?
-
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
S-adenosyl-L-methionine + 7Me-GpppA-C5
S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
show the reaction diagram
S-adenosyl-L-methionine + 7Me-GpppA-Cn
S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-Cn
show the reaction diagram
-
exclusive methylation at the 2'O position
-
-
?
S-adenosyl-L-methionine + GpppA-Cn
S-adenosyl-L-homocysteine + GpppA-2'OMe-Cn
show the reaction diagram
-
exclusive methylation at the 2'O position
-
-
?
S-adenosyl-L-methionine + m7G(5')pppR-RNA
S-adenosyl-L-homocysteine + m7G(5')pppRm-RNA
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
-
110% activity in the presence of 10 mM KCl, in Tris-HCl buffer (pH 7.5)
KCl
-
activation, 140-180 mM, cap I-methyltransferase
additional information
-
no divalent cations required
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3'-deoxy-ribavirin 5'-triphosphate
binds almost as efficiently as ribavirin 5-triphosphate
5',N8-Adenosyl-alpha,beta-diaminobutyric acid
-
moderate
5'-S-isobutylthio-5'-deoxyadenosine
-
about 25% inhibition at 0.1 mM
5-ethynyl-1-beta-D-ribofuranosylimidazole-4-carboxamide triphosphate
-
about 40% inhibition at 0.1 mM
7-methylguanosine(5')pppN-dinucleotides
-
weak, competitive
A9145C
acyclovir triphosphate
-
deazaadenosine
-
about 5% inhibition at 0.1 mM
EICAR 5'-triphosphate
-
Mg2+
-
almost complete inhibition at 100 mM
NaCl
-
above 0.05 M
ribavirin
-
about 10% inhibition at 0.1 mM
ribavirin 5'-triphosphate
50% inhibition at 0.1 mM
ribavirin triphosphate
-
about 50% inhibition at 0.1 mM
S-(2-azaadenosyl)-L-homocysteine
-
-
S-(3'-aminoadenosyl)-L-homocysteine
-
-
-
S-(3-deazaadenosyl)-L-homocysteine
-
strong
S-(5'-adenosyl)-L-cysteine
-
about 25% inhibition at 0.1 mM
S-(8-azaadenosyl)-L-homocysteine
-
weak
S-(N6-Dimethyl-3-deazaadenosyl)-L-homocysteine
-
weak
S-(N6-Methyladenosyl)-L-homocysteine
-
-
S-5'-adenosyl-L-cysteine
-
about 5% inhibition at 0.1 mM
S-Adenosyl-D-homocysteine
-
-
S-adenosyl-L-cysteine
-
moderate
S-adenosyl-L-homocysteine
S-Adenosyl-L-homocysteine sulfone
-
strong
S-adenosyl-L-homocysteine sulfoxide
-
strong
S-Aristeromycinyl-L-homocysteine
-
-
S-Cytidyl-L-homocysteine
-
weak
S-Inosyl-L-homocysteine
-
weak
S-Tubercidinyl-L-homocysteine
-
strong
S-Uridyl-L-homocysteine
-
weak
sinefungin
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2',3',5'-tri-O-acetyladenosine
-
about 50% increased activity at 0.1 mM
3-deazaadenosine
-
about 10% increased activity at 0.1 mM
5'-deoxy-5'-(methylthio)adenosine
-
about 75% increased activity at 0.1 mM
5'-deoxy-5'-methylthioadenosine
-
about 10% increased activity at 0.1 mM
5'-S-isobutylthio-5'-deoxyadenosine
-
about 60% increased activity at 0.1 mM
7,2',3',5'-tri-O-acetyladenosine
-
about 20% increased activity at 0.1 mM
-
nsp10 protein
-
ribavirin
-
about 10% increased activity at 0.1 mM
ribavirin triphosphate
-
about 20% increased activity at 0.1 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000394
7Me-GpppA-C5
-
in 40 mM Tris-HCl, pH 7.5, at 30C
0.000005
brome mosaic virus RNA
-
-
-
0.0000014
G(5')pppA-poly(A)
-
-
0.0000016
m7G(5')pppA-poly(A)
-
-
1.8
m7G(5')pppG-RNA
-
apparent value, in glycine buffer (pH 10.0), at 22C
-
0.00016 - 0.00023
RNA chain length 2-6 nt
-
-
-
0.000005 - 0.000015
RNA chain length 20-50 nt
-
-
-
0.0006 - 0.19
S-adenosyl-L-methionine
additional information
additional information
-
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000018 - 0.00000134
A9145C
0.0046
S-(3'-aminoadenosyl)-L-homocysteine
-
-
-
0.0021
S-(3-deazaadenosyl)-L-homocysteine
-
-
0.009
S-(N6-Methyladenosyl)-L-homocysteine
-
-
0.014
S-Adenosyl-D-homocysteine
-
-
0.058
S-adenosyl-L-cysteine
-
-
0.00053 - 0.001
S-adenosyl-L-homocysteine
0.0012
S-Adenosyl-L-homocysteine sulfone
0.0026
S-Aristeromycinyl-L-homocysteine
-
-
0.0012
S-Tubercidinyl-L-homocysteine
-
-
0.0000075 - 0.0000194
sinefungin
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00034 - 0.144
S-adenosyl-L-homocysteine
0.000041 - 0.00063
sinefungin
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00000233
-
purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
about pH 7.0
additional information
-
pI: 8.4
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9
-
-
6.5 - 8
-
m7G- and A-capped RNA substrates
6.6 - 8.1
-
about half-maximal activity at pH 6.6 and 8.1
7 - 9
-
-
additional information
-
beginning from pH 6.9 continous increase of activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
cap I- and cap II-methyltransferase
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
-
sucrose density gradient centrifugation
38000
-
gel filtration
65000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
2 * 35000, SDS-PAGE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
to 2.5 A resolution. Residues 175 to 377 encode a distinct 2'-O MT domain segregated from the other domains and responsible for methylation of cap0 structure
hanging drop method at room temperature, 0.1 M sodium citrate pH 5.8, 1.2 M lithium sulfate, 0.5 M ammonium sulfate, vapour diffusion for 1 week, structure analysis with and without bound nucleotide, model
in complex with ribavirin 5-triphosphate and S-adenosyl-L-methionine
by vapour-diffusion, to 2.8 A resolution, in complex with S-adenosyl-l-methionine, crystals belong to the tetragonal space group P43, with unit-cell parameters a = b = 83.0, c = 170.2 A, presence of four MTase molecules per asymmetric unit and a corresponding solvent content of 49.9%, residues Lys62, Asp147, Lys184 and Glu220 specifically characterize the active-site region
-
hanging drop vapor diffusion method, using in 0.1 M MES, pH 5.0, 2 M NaCl, 0.1 M NaH2PO4, 0.1 M KH2PO4 after 24 h at 25C
by vapour-diffusion, to 2.7 A resolution, in complex with S-adenosyl-l-methionine, crystals belong to the orthorhombic space group P21212, with unit-cell parameters a = 58.7, b = 126.6, c = 156.1 A, presence of three MTase molecules in the asymmetric unit, with a solvent content of 57.7%, residues Lys62, Asp147, Lys184 and Glu220 specifically characterize the active-site region
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
binding site of the enzyme for RNA, immediately downstream, is pH-sensitive, above pH 7.5: dramatic increase of Km
639484
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
10 min stable
44
-
and above, inactivation after 20 min
50
-
and below, stable in the presence of 10% v/v glycerol and 250 mg/l bovine serum albumin
59
-
rapid inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
bovine serum albumin stabilizes
-
glycerol, 10% v/v stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, several months
-
4C, several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography on ADP-Sepharose
-
by centrifugation
-
by gel filtration
fragment mutant and exchange mutants from recombinant E. coli
-
GSTPrep 16/10 column chromatography
-
HeLa-cells, phosphocellulose chromatography
-
HisPur cobalt resin column chromatography
-
mutants recombinant from E. coli
-
Ni-NTA resin column chromatography
-
Ni-NTA resin column chromatography and Superdex 200 gel filtration
nickel-nitrilotriacetic acid resin column chromatography and heparin-Sepharose 6 column chromatography
-
recombinant from Spodoptera frugiperda cells
-
recombinant His-tagged N-terminal part of NS5 protein from E. coli
Sepharose resin column chromatography and Superdex 200 gel filtration
-
solubilized from viral core
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
as N-terminal His6-tag expressed in a pDEST 14 HN expression vector and transformed into Escherichia coli Rosetta (DE3) pRos
baculovirus expression system
cloning of His-tagged N-terminal part of NS5 protein, termed NS5MTaseDV, amino acids 1-296, overexpression in Escherichia coli
expressed in Escherichia coli BL21 cells
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli C41 (DE3) cells
-
expressed in Escherichia coli C41(DE3) cells
-
expression of VP4 protein in Spodoptera frugiperda cells
-
expression of wild-type, exchange mutants and open-reading-frame mutant in Escherichia coli, all glutathione-S-transferase-tagged
-
mutant C272S/K175C/R209K
mutant enzymes are expressed in Escherichia coli and the tag is cleaved off
-
transfection of BHK-21 cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D265E
complete loss of 2'-O-methylation activity, Bluetongue virus replication in cells is abrogated
D265V
complete loss of 2'-O-methylation activity, Bluetongue virus replication in cells is abrogated
N311A
2fold reduction of 2'-O-methylation activity, Bluetongue virus replication in cells is abrogated
N311A/Y334/R367A
2fold reduction of 2'-O-methylation activity, Bluetongue virus replication in cells is abrogated
R367A
mutation does not affect methyl group transfer
Y334A
mutation does not affect methyl group transfer
D113A
-
about 110% substrate binding and activity compared to the wild type enzyme
D129A
-
about 40% substrate binding and activity compared to the wild type enzyme
D221A
-
about 115% substrate binding and activity compared to the wild type enzyme
D247A
-
about 75% substrate binding and activity compared to the wild type enzyme
E202A
-
about 30% substrate binding and activity compared to the wild type enzyme
F173A
-
about 20% substrate binding and activity compared to the wild type enzyme
K169A
-
about 20% substrate binding and activity compared to the wild type enzyme
K45A
-
about 1% substrate binding and activity compared to the wild type enzyme
W4A
-
about 90% substrate binding and activity compared to the wild type enzyme
Y14A
-
about 10% substrate binding and activity compared to the wild type enzyme
Y29A
-
about 15% substrate binding and activity compared to the wild type enzyme
K239A
-
the mutant shows no activity
D106A
-
the mutant shows 38% of wild type activity
D130A
-
the mutant shows 2% of wild type activity
D99A
-
inactive
E203A
-
inactive
G73A
-
the mutant shows 18% of wild type activity
G94A
-
the nsp10 mutation binds nsp16 with a slightly reduced affinity (60%) but is still able to stimulate nsp16 2'O-MTase activity
G94D
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
H83A/P84A
inactive
I40A
-
the mutant shows 8% of wild type activity
K170A
-
inactive
K46A
-
the mutant shows 1% of wild type activity
K93E
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
K95A
-
the nsp10 mutation binds nsp16 with a slightly reduced affinity (80%) but is still able to stimulate nsp16 2'O-MTase activity
L244A
-
inactive
M247A
-
inactive
M41A
-
the mutant shows 4% of wild type activity
M44A
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
N43A
-
the mutant shows 11% of wild type activity
Q87A
-
the mutant shows 62% of wild type activity
R78A
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
R78G
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
R86A
-
inactive
S188A
-
the substitution shows 72% loss of activity with no significant effect on the stability of the nsp10/nsp16 complex
S72A
-
the mutant protein binds nsp16 but weakly activates 2'O-MTase activity
T48A
-
the mutant shows 20% of wild type activity
T54E
-
the substitution shows 72% loss of activity with no significant effect on the stability of the nsp10/nsp16 complex
T58A
-
the substitution shows 43% loss of activity with no significant effect on the stability of the nsp10/nsp16 complex
T58E
-
the substitution shows 99% loss of activity with 54% association of the Nsp10/nsp16 complex compared to the wild type
T58N
-
the substitution shows 70% loss of activity with no significant effect on the stability of the nsp10/nsp16 complex
V104G
-
the mutant shows 4% of wild type activity
V42A
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
V44A
-
the mutant shows 1% of wild type activity
V78A
-
inactive
Y132A
-
the mutant shows 5% of wild type activity
Y132F
-
the mutant shows 9% of wild type activity
Y132H
-
inactive
Y132T
-
the mutant shows 5% of wild type activity
Y30A
-
the mutant shows 1% of wild type activity
Y30F
-
the mutant shows 6% of wild type activity
Y76A/C77A/R78A
inactive
Y96I
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
Y96V
-
the nsp10 mutation abrogates stimulation of nsp16 enzyme activity
G70A
-
the mutant shows reduced activity compared to the wild type
-
H83A/P84A
-
inactive
-
K93A
-
the mutant shows reduced activity compared to the wild type
-
Y76A/C77A/R78A
-
inactive
-
K266A
proteins similar to that of the wild-type TbMT57-HA protein, defective in the biogenesis of the SL cap 4 structure
C178S
-
point mutations, glutathione-S-transferase-tagged, exchange shows no effect
C272S
-
point mutations, glutathione-S-transferase-tagged, exchange shows no effect
C272S/K175C/R209K
2-bromoethylamine reveals 64% target site modification in a overnight reaction at 37C, unmodified mutant has no detectable methyltransferase activity, whereas the modified protein is active, exhibiting 20-30% of the specific catalytic rate of wild-type
D182A
-
m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
D182A/E233A
-
m7G binding pocket double mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
E233A
-
m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
F180A
-
m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
F180W
-
no apparent defects in catalytic activity
G96D
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double mutant G96D plus Cts23 mutation in gene A18 shows that the J3-mutants serves as an extragenic suppressor of A18-Cts23 mutant; mutation in J3 protein with nucleoside-2-O'-methyltransferase activity evokes a phenotype with abnormally long RNA transcripts analogously to A18 gene-mutation Cts23
K175C
aziridine reveals 66.7% target site modification in a 4 h, room temperature reaction, equivalent modification level achieved in an overnight incubation using 2-bromoethylamine at 37C, but protein losses due to aggregation during the modification reaction are, notably, negligible in the lower temperature/shorter duration reactions employed for aziridine
Y22A
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m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
D1762A
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with pinpoint plaque morphologies and replication defects in single-step growth assays, viral RNA and protein synthesis is diminished
E1674A
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the mutation in the V1 region of L protein abolishes 2'-O methylation activity
E1674D
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the mutation in the V1 region of L protein shows severely reduced 2'-O methylation activity (about 20%) compared to the wild type
E1674K
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the mutation in the V1 region of L protein shows severely reduced 2'-O methylation activity (about 15%) compared to the wild type
E1674Q
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the mutation in the V1 region of L protein shows severely reduced 2'-O methylation activity (about 15%) compared to the wild type
E1833A
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with pinpoint plaque morphologies and most significant replication defects in single-step growth assays, viral RNA and protein synthesis is diminished
E1833Q
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delay in replication
F1691A
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the mutation in the V1 region of L protein abolishes 2'-O methylation activity
F1691W
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the mutation in the V1 region of L protein shows slightly reduced 2'-O methylation activity (about 78%) compared to the wild type
F1691Y
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the mutation in the V1 region of L protein shows slightly reduced 2'-O methylation activity (about 80%) compared to the wild type
G1674A
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does not significantly perturb viral growth and gene expression, replicates with almost indistinguishable kinetics to recombinant vesicular stomatitis virus
K1651A
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with pinpoint plaque morphologies and most significant replication defects in single-step growth assays, viral RNA and protein synthesis is diminished
K1795A
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most significant defect in replication; with pinpoint plaque morphologies and replication defects in single-step growth assays, viral RNA and protein synthesis is diminished
Y1650A
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the mutation in the V1 region of L protein abolishes 2'-O methylation activity
Y1650F
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the mutation in the V1 region of L protein shows slightly reduced 2'-O methylation activity (about 85%) compared to the wild type
Y1650W
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the mutation in the V1 region of L protein shows slightly reduced 2'-O methylation activity (about 90%) compared to the wild type
E218A
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the mutation abolishes 2'-O-methyltransferase activity and enhances replication but not virulence in Ifit1-deficient mice after peripheral infection
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
medicine
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the nsp10-nsp16 interface represents an attractive target for antivirals against human and animal pathogenic coronaviruses
additional information
Show AA Sequence (178 entries)
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