Information on EC 2.1.1.56 - mRNA (guanine-N7)-methyltransferase

Word Map on EC 2.1.1.56
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.56
-
RECOMMENDED NAME
GeneOntology No.
mRNA (guanine-N7)-methyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + a 5'-(5'-triphosphoguanosine)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N7-methyl 5'-triphosphoguanosine)-[mRNA]
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
methyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
mRNA capping I
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:mRNA (guanine-N7)-methyltransferase
The nucleoside next to the terminal guanosine may be either guanosine or adenosine.
CAS REGISTRY NUMBER
COMMENTARY hide
56941-25-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
activity belongs to the N-terminal part of the replicase
-
-
Manually annotated by BRENDA team
enzyme activity belongs to VP4 protein
-
-
Manually annotated by BRENDA team
L929-cells
-
-
Manually annotated by BRENDA team
mutant met-
-
-
Manually annotated by BRENDA team
strain VY1160, osmotic sensitive
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain WR, from infected HeLa cells
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
mutation defective in N-7 methylation but not 2-O'-methylation is lethal for DNV replication
physiological function
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 + dGTP
S-adenosyl-L-homocysteine + m7dGTP
show the reaction diagram
S-adenosyl-L-methionine + G(5')pppA
S-adenosyl-L-homocysteine + m7G(5')pppA
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + G(5')pppG
S-adenosyl-L-homocysteine + m7G(5')pppG
show the reaction diagram
-
and dGTP, best substrates
-
-
?
S-adenosyl-L-methionine + G(5')pppN
S-adenosyl-L-homocysteine + m7G(5')pppG
show the reaction diagram
S-adenosyl-L-methionine + G(5')pppR-RNA
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
show the reaction diagram
S-adenosyl-L-methionine + GDP
S-adenosyl-L-homocysteine + m7GDP
show the reaction diagram
-
-
-
-
S-adenosyl-L-methionine + GpppA
S-adenosyl-L-homocysteine + m7GpppA
show the reaction diagram
S-adenosyl-L-methionine + GpppA-RNA
S-adenosyl-L-homocysteine + m7GpppA-RNA
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + GpppAC3
S-adenosyl-L-homocysteine + m7GpppAC3
show the reaction diagram
-
is efficiently methylated at the guanine-N7 position
-
-
?
S-adenosyl-L-methionine + GpppG
S-adenosyl-L-homocysteine + m7GpppG
show the reaction diagram
S-adenosyl-L-methionine + GTP
S-adenosyl-L-homocysteine + m7GTP
show the reaction diagram
S-adenosyl-L-methionine + guanylimidodiphosphate
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + m7G(5')pppG
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + pp(A)n
?
show the reaction diagram
-
-
-
-
?
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 + G(5')pppR-RNA
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
-
-
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate
-
i.e. CHAPS, strong inhibition
5',N8-Adenosyl-alpha,beta-diaminobutyric acid
-
moderate
amino acid modified structure analogue of adenosyl-L-homocysteine
-
i.e. A9145C, strong
-
aurintricarboxylic acid
aza-S-adenosyl-L-methionine
-
-
carbocyclic aza-S-adenosyl-L-methionine
-
-
Cu2+
-
strong
cycloheximide
-
inhibits protein synthesis in infected BHK cells
deoxycholate
-
inactivation
Digitonin
-
strong inhibition
GpppG
-
inhibits binding of the enzyme to RNA
N,N-bis-(3-D-gluconamidopropyl)-deoxycholamide
-
strong inhibition
n-octyl-beta-D-gluconpyranoside
-
i.e. octylglucoside, strong inhibition
N-tetradecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate
-
i.e. zwittergent 3-14, strong inhibition
Oligoadenylic acid mono- and triphosphates
-
S-(2-azaadenosyl)-L-homocysteine
-
moderate
S-(3-Aminoadenosyl)-L-homocysteine
-
weak
S-(3-deazaadenosyl)-L-homocysteine
S-(8-azaadenosyl)-L-homocysteine
S-(N6-Dimethyl-3-deazaadenosyl)-L-homocysteine
-
weak
S-(N6-Methyladenosyl)-L-homocysteine
S-Adenosyl-D-homocysteine
S-adenosyl-homocysteine
-
-
S-adenosyl-L-cysteine
S-adenosyl-L-homocysteine
S-Adenosyl-L-homocysteine structural analogues
S-Adenosyl-L-homocysteine sulfone
S-adenosyl-L-homocysteine sulfoxide
S-Aristeromycinyl-L-homocysteine
S-Inosyl-L-homocysteine
-
weak
S-Tubercidinyl-L-homocysteine
sinefungin
Thesit
-
strong inhibition
-
Triton X-100
-
inactivation, reversible by addition of lipids: cardiolipin, phosphatidylglycerol, and especially phosphatidylserine
Zn2+
-
strong
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
importin-alpha
-
enhances specific binding of the enzyme to RNA, enhances RNA modification activity in vivo and in vitro
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.085 - 0.101
G(5')pppG
0.023 - 1.8
G(5')pppR-RNA
0.1 - 0.24
GpppA
0.033 - 1
GTP
0.002 - 0.18
S-adenosyl-L-methionine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0001947 - 0.022
G(5')pppR-RNA
0.0983
GpppA
Saccharomyces cerevisiae
-
-
0.000054 - 0.022
S-adenosyl-L-methionine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000018 - 0.00000134
amino acid modified structure analogue of adenosyl-L-homocysteine
-
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
-
-
0.0012
S-adenosyl-L-homocysteine sulfoxide
-
-
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
0.0023 - 0.0042
aurintricarboxylic acid
0.1
aza-S-adenosyl-L-methionine
Encephalitozoon cuniculi
-
-
0.035
carbocyclic aza-S-adenosyl-L-methionine
Encephalitozoon cuniculi
-
-
0.00177
S-adenosyl-homocysteine
Dengue virus
-
pH 7.5, 20C
0.004 - 0.021
S-adenosyl-L-homocysteine
0.000021 - 0.0015
sinefungin
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00000235
-
purified enzyme
0.000593
-
purified enzyme
0.0196
-
purified D1 fragment, residues 498-844
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2 - 7.8
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
-
half-maximal activity at pH 5.0, unstable above pH 8.0
6 - 7.5
-
methyl transfer by Ecm1 declines sharply below pH 6.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
19 - 37
-
-
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
70% of total activity, 1/3 of which is losely associated to ribosomes
Manually annotated by BRENDA team
-
lipid-binding site, membrane binding activity of wild-type and diverse mutants
Manually annotated by BRENDA team
-
of infected BHK cells
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Dengue virus type 2 (strain Puerto Rico/PR159-S1/1969)
Dengue virus type 2 (strain Thailand/NGS-C/1944)
Encephalitozoon cuniculi (strain GB-M1)
Encephalitozoon cuniculi (strain GB-M1)
Encephalitozoon cuniculi (strain GB-M1)
Encephalitozoon cuniculi (strain GB-M1)
Encephalitozoon cuniculi (strain GB-M1)
Encephalitozoon cuniculi (strain GB-M1)
Encephalitozoon cuniculi (strain GB-M1)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vaccinia virus (strain Western Reserve)
Vesicular stomatitis Indiana virus (strain Mudd-Summers)
Yellow fever virus (strain 17D vaccine)
Yellow fever virus (strain 17D vaccine)
Yellow fever virus (strain 17D vaccine)
Yellow fever virus (strain 17D vaccine)
Yellow fever virus (strain 17D vaccine)
Yellow fever virus (strain 17D vaccine)
Zika virus (strain Mr 766)
Zika virus (strain Mr 766)
Zika virus (strain Mr 766)
Zika virus (strain Mr 766)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30000
-
SDS-PAGE
31000
-
1 * 31000 + 1 * 95000, SDS-PAGE
33000
-
small subunit of mRNA capping enzyme , gel filtration, SDS-PAGE
39000
-
x * 39000, SDS-PAGE of His-tagged TbCmt1
40000
-
1 * 40000, SDS-PAGE
47000
Western blot
49000
-
gel filtration
56000
-
gel filtration, sucrose density gradient centrifugation
58660 - 60000
DNA sequence determination, western blot
68820 - 70000
DNA sequence determination, western blot
95000
-
1 * 31000 + 1 * 95000, SDS-PAGE
97000
-
large subunit of mRNA capping enzyme, gel filtration, SDS-PAGE
120000 - 129000
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 39000, SDS-PAGE of His-tagged TbCmt1
dimer
-
1 * 31000 + 1 * 95000, SDS-PAGE
heterodimer
-
the guanine-N7 methyltransferase domain of vaccinia virus mRNA capping enzyme is a heterodimer composed of a catalytic subunit and a stimulatory subunit, X-ray crystallography
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
6.5 mg/ml purified native or selenomethionine-labeled, or Hg-labeled enzyme in complex with S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, or the cap guanylate plus S-adenosyl-L-homocysteine, in 20 mM Tris-HCl, pH 8.0, 220 mM NaCl, vapour diffusion against a well solution of 1.2 M sodium potassium tartrate, 50 mM bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, pH 6.0-6.2, and 20 mM DTT, X-ray diffraction structure determination and analysis at 2.0-2.7 A resolution, modeling
-
Ecm1-aza-S-adenosyl-L-methionine binary complex, by vapor diffusion, reveals that the inhibitor occupies the same site as S-adenosyl-L-methionine
-
Ecm1-sinefungin binary complex, by vapor diffusion, to 2.6 A resolution
-
structure-function relationships of the N7-MTase. The ExoN domain is closely involved in the activity of the N7-MTase. Two of the 12 critical residues essential for the N7-MTase are located at the N terminus of the core ExoN domain, reinforcing a role of the ExoN domain in the N7-MTase activity of nsp14. The other 10 critical residues are distributed throughout the N7-MTase domain but localized mainly in the S-adenosyl-L-methionine-binding pocket
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
inactivation after 16 h at 0C
485464
7.5
-
16 h stable at 0C
485464
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
cardiolipin, phosphatidylglycerol, and phosphatidylserine
-
expression and enzyme activity stable for 3 h in infected BHK cells
-
gel electrophoresis inactivates
-
small and large subunit of the mRNA capping enzyme are not stable and active when isolated, respectively
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, in 10% v/v glycerol, at least several months
-
-20C, Tris buffer, pH 7.5, 50% glycerol
-
-80 C, Tris-HCl buffer, pH 7.5, 10% glycerol
-
-80C, several months
0C, 16 h at pH 7.5
-
4C or -20C, several months, stable
-
4C, t1/2: less than 36 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by immobilized-metal-affinity chromatography
-
by nickel-affinity chromatography
-
co-purifies with mRNA-guanylyltransferase
-
DNA-agarose affinity chromatography
enzyme and truncated mutants recombinant from Saccharomyces cerevisiae
-
from infected HeLa cells
-
His6-Smt3-Abd1 by nickel-nitrilotriacetic acid fast flow column chromatography, Abd1 free of the tag by gel filtration, to homogeneity
-
methyltransferase domain, and small subunit of mRNA capping enzyme, recombinant from E. coli
-
Ni2+-NTA agarose column chromatography
-
recombinant Ecm1 mutant proteins, by nickel-agarose chromatography
-
recombinant from E. coli and recombinant from in vitro transcription and translation
-
recombinant from Spodoptera frugiperda cells
-
recombinant N-terminally His6-tagged enzyme as Smt3-fusion protein from Escherichia coli strain BL21(DE3), removal of His-Smt3 tag by specific protease Ulp1, purification by metal affinity chromatography and gel filtration
-
small and large subunit of mRNA capping enzyme , recombinant from E. coli
-
TbCmt1 protein purified from soluble extract by nickel-agarose, followed by phosphocellulose chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 carboxyl-truncated versions of the large subunit of the mRNA capping enzyme, expression in Escherichia coli; coexpression of genes D1 and D12 from 1 plasmid; vaccinia virus genes D1 and D12 encoding the large and the small subunit of the mRNA capping enzyme, respectively, individual overexpression in Escherichia coli BL21
-
2 carboxyl-truncated versions of the large subunit of the mRNA capping enzyme, expression in Escherichia coli; vaccinia virus genes D1 and D12 encoding the large and the small subunit of the mRNA capping enzyme, respectively, individual overexpression in Escherichia coli BL21
-
as a His6-Smt3 fusion protein in Escherichia coli BL21(DE3) CodonPlus RIL using a pET-based pSmt3-TOPO vector
-
complementation of knock-down HeLa S3 cells with enzymatically active N-truncated enzyme comprising residues 120-476 and being located at the nucleus, no complementation of knock-down HeLa S3 cells with enzymatically active N-truncated enzyme comprising residues 144-476 and being loacetd mainly in the cytoplasm, truncated enzymes are expresss in fusion with GFP
-
D1 fragment, amino acids 498 to 844; expression of D1 fragment mutants in Escherichia coli; individual and coexpression of His-tagged D1 large subunit fragment, being the methyltransferase domain of the enzyme, and of D12 small subunit in Escherichia coli
-
D1 fragment, amino acids 498 to 844; individual and coexpression of His-tagged D1 large subunit fragment, being the methyltransferase domain of the enzyme, and of D12 small subunit in Escherichia coli
-
D1 fragment, amino acids 520-844; individual and coexpression of His-tagged D1 large subunit fragment, being the methyltransferase domain of the enzyme, and of D12 small subunit in Escherichia coli
-
expressed in Saccharomyces cerevisiae strains YBS40, YBS2, and YBS20
-
expression in Escherichia coli, His-tagged; expression in Escherichia coli, His-tagged, amino acid sequence analysis; expression in Escherichia coli, His-tagged, amino acid sequence analysis
expression in Escherichia coli; in vitro transcription and translation in presence of liposomes, enzyme is attached to the liposomes surface
-
expression in Saccharomyces cerevisiae, His-tagged, amino acid sequence analysis
expression of 2 different variants, each with and without His-tag, in Escherichia coli, amino acid sequence analysis
-
expression of full length and 2 truncated genes in Saccharomyces cerevisiae and Escherichia coli, location of methyltransferase activity
-
expression of full length enzyme and truncated enzyme in yeast and in HeLa cells, His-tagged, as GFP-fusion protein, or cMyc-tagged
-
expression of N-terminally His6-tagged enzyme, residues 1-298, as Smt3-fusion protein in Escherichia coli strain BL21(DE3) or strain B834DE3, expression of wild-type and mutant alleles under control of a yeast promotor in a Saccharomyces cerevisiae abd1DELTA null mutant strain
-
expression of VP4 protein in Spodoptera frugiperda cells
-
His6-tagged proteins are expressed in Escherichia coli BL21(DE3) cells and in Saccharomyces cerevisiae strain YBS40
-
inserted into vector pET16b and transformed into Escherichia coli BL21(DE3), yeast strain YBS40 transformed with CEN TRP1 plasmids containing the wild-type and mutant alleles of ECM1
-
into vector pDest14 and transformed into Escherichia coli Rosetta(DE3) cells
-
TbCmt1 inserted into an inducible T7 RNA polymerase-based pET vector and introduced into Escherichia coli BL21(DE3)Gold
-
the MTase domain, representing the N-terminal 272 amino acids of DENV-4 NS5 is cloned and purified
-
vaccinia virus genes D1 and D12 encoding the large and the small subunit of the mRNA capping enzyme, respectively, individual overexpression in Escherichia coli BL21
-
wild-type or mutant vD12, into the pRS413T vector, expressed in Saccharomyces cerevisiae strain YBS40
-
yeast strain YBS40 transformed with CEN TRP1 plasmids containing the wild-type and mutant alleles of ECM1, the pET16-Ecm1 plasmids introduced into Escherichia coli BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D146A
-
mutant completely loses the N-7 activity (methylation of N-7 position of guanine) and 2'-O methylation activity. Mutation defective in N-7 methylation is lethal for DNV replication
E217A
-
mutants retains 59% of wild-type N-7 (methylation of N-7 position of guanine) activity
K181A
-
mutants retains 16% of wild-type N-7 (methylation of N-7 position of guanine) activity
K61A
-
mutants retains 34% of wild-type N-7 (methylation of N-7 position of guanine) activity
D122A/S123A
-
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, only slightly at 37C
E225D
-
growth at all temperatures
E225Q
-
growth at all temperatures
F141H
-
grows well at 25 and 30C but forms pinpoint colonies at 37C
F141I
-
grows well at 25 and 30C but forms pinpoint colonies at 37C
F141L
-
grows as well as the wild-type ECM1 strain at all temperatures
F141V
-
grows well at 25 and 30C but forms pinpoint colonies at 37C
F214L
-
growth at all temperatures, slow growth at 37C
H144A/Y145F
-
fully functional
H144A/Y145L
-
defective in vivo, fails to grow at 37C, forms microcolonies at 30C, and grows slowly at 25C
I95A
-
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C, 30C, and 37C, no effect on enzyme function
I95A/Y124A
-
site-directed mutagenesis, only slight complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, growth defects
K75Q
-
slow growth at all temperatures
K75R
-
growth at all temperatures
L216A
-
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C, no effect on cell growth
N50A
-
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C, only modest growth defects
N50A/Y284A
-
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
P175A
-
grows as well as the wild-type ECM1 strain at all temperatures
R47A
-
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C
R47A/K75A
-
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Y124F
-
growth at all temperatures, slow growth at 37C
Y145H
-
grows as well as the wild-type ECM1 strain at all temperatures
Y145I
-
intermediate phenotype of slow growth at 25 and 30C and microcolony formation at 37C
Y212F
-
growth at all temperatures
Y284A
-
site-directed mutagenesis, some complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, slow growth phenotype
C416R
-
mutation abolishes the N7-MTase activity on GTP
D273A
-
97% of wild-type methylation activity
D331E
-
70% of wild-type methylation activity
D90A/E92A
G416R
-
45% of wild-type methylation activity
K61A
-
97% of wild-type methylation activity
L419A
-
complete loss of methylation activity
L419R
-
15-50% of wild-type activity
N256A
-
97% of wild-type methylation activity
N334A
-
89% of wild-type methylation activity
N388A
-
83% of wild-type methylation activity
D144A
-
TbCmt1 mutant shows 5% of wild-type activity
D80A
-
TbCmt1 mutant is unable to support methylation at levels of input protein sufficient to convert all the input substrate to the methylated form by wild-type TbCmt1, specific activity is less than 0.1% of the wild-type activity
E258A
-
TbCmt1 mutant is unable to support methylation at levels of input protein sufficient to convert all the input substrate to the methylated form by wild-type TbCmt1, specific activity is less than 0.1% of the wild-type activity
F171A
-
TbCmt1 mutant displays near-wild-type activity
H174A
-
TbCmt1 mutant displays near-wild-type activity
K83A
-
TbCmt1 mutant is unable to support methylation at levels of input protein sufficient to convert all the input substrate to the methylated form by wild-type TbCmt1, specific activity is less than 0.1% of the wild-type activity
V241A
-
TbCmt1 mutant displays near-wild-type activity
V242A
-
TbCmt1 mutant displays near-wild-type activity
Y175A
-
TbCmt1 mutant shows 3% of wild-type activity
Y246A
-
TbCmt1 mutant shows 2% of wild-type activity
C173A/S174A
-
modest defect, grows well at 19, 30 and 34C, but very slowly at 37C
D192A/S193A
-
temperature-sensitive vD12 allele, near-lethality, only grows very slowly at 19C
D52A/L53A
-
modest defect, grows well at 30C, but slowly at 34C and not at 37C
D545A
-
the mutant has 102% of wild type activity
D598E
-
the mutant enzyme has 1% of wild type activity
D598N
-
the mutant enzyme hasless than 1% of wild type activity
D604E
-
the mutant enzyme has 112% of wild type activity
D604N
-
the mutant enzyme has less than 1% of wild type activity
D620E
-
the mutant enzyme has 59% of wild type activity
D620N
-
the mutant enzyme has 11% of wild type activity
D657A
-
the mutant enzyme has 112% of wild type activity
D676A
-
the mutant enzyme has less than 1% of wild type activity
D676E
-
the mutant enzyme has 40% of wild type activity
D676N
-
the mutant enzyme has less than 1% of wild type activity
E104A/G11A
-
modest defect, grows well at 19, 30 and 34 C, but slowly at 37C
E24A-25A
-
temperature-sensitive vD12 allele, does not grow at 37C
E275A/N276A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
E763A
-
the mutant enzyme has 16% of wild type activity
E763D
-
the mutant enzyme has 1% of wild type activity
E763Q
-
the mutant enzyme has 32% of wild type activity
E816A
-
the mutant enzyme has 76% of wild type activity
F141A/R142A
-
grows slowly at 19 and 30C and fails to grow at 34C or higher; temperature-sensitive vD12 allele, does not grow above 34C
F176A/K177A
-
lethal at all temperatures
F44A/L45A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
F556A
-
the mutant has 1% of wild type activity
F556L
-
the mutant has 3% of wild type activity
F609A
-
the mutant enzyme has 117% of wild type activity
F659A
-
the mutant enzyme has 96% of wild type activity
F679A
-
the mutant enzyme has 5% of wild type activity
F679H
-
the mutant enzyme has 13% of wild type activity
F679I
-
the mutant enzyme has 18% of wild type activity
F679L
-
the mutant enzyme has 55% of wild type activity
F679N
-
the mutant enzyme has 18% of wild type activity
F679V
-
the mutant enzyme has 20% of wild type activity
F814A
-
the mutant enzyme has 99% of wild type activity
F815A
-
the mutant enzyme has 92% of wild type activity
G600A
-
in D1 fragment, amino acids 498-844, 4% activity remaining
H260A/S261A
-
modest defect, grows well at 19, 30 and 34C, but very slowly at 37C
H682N
-
the mutant enzyme has 1% of wild type activity
H682Q
-
the mutant enzyme has 4% of wild type activity
I47A/S48A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
K111A/R112A
-
modest defect, grows well at 19, 30 and 34C, but very slowly at 37C
K156A
-
viable at 19C and 30C, but fails to grow at 34C
K156A/L157A
-
lethal at all temperatures
K223A/D223A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
K573A
-
the mutant enzyme has less than 1% of wild type activity
K573Q
-
the mutant enzyme has less than 1% of wild type activity
K573R
-
the mutant enzyme has 1% of wild type activity
K607A
-
the mutant enzyme has101% of wild type activity
K607Q
-
the mutant enzyme has 74% of wild type activity
K607R
-
the mutant enzyme has 13% of wild type activity
L167A
-
viable at 19C and 30C, but fails to grow at 34C
L16A/P17A
-
temperature-sensitive vD12 allele, forms small colonies at 19C and fails to grow at 30C or higher
N120A/N121A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
N126A/N127A
-
modest defect, grows well at 19, 30 and 34C, but very slowly at 37C
N42A-Y43A
-
temperature-sensitive vD12 allele, does not grow at 37C
N54A/R55A
-
modest defect, grows well at 19 and 30C, but slowly at 34 and very slowly at 37C
N550A
-
the mutant has 3% of wild type activity
N550D
-
the mutant has 19% of wild type activity
N550Q
-
the mutant has 10% of wild type activity
N570A
-
the mutant enzyme has 37% of wild type activity
N570D
-
the mutant enzyme has 21% of wild type activity
N570Q
-
the mutant enzyme has 73% of wild type activity
N601A
-
the mutant enzyme has 32% of wild type activity
P158A/T159A
-
temperature-sensitive vD12 allele, only grows at 19C
P35A/S36A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
Q678A
-
the mutant enzyme has 55% of wild type activity
R280A-R281A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
R548A
-
the mutant has 59% of wild type activity
R560A
-
the mutant has 18% of wild type activity
R560K
-
the mutant has 84% of wild type activity
R560Q
-
the mutant has 84% of wild type activity
R562A
-
the mutant has 10% of wild type activity
R562K
-
the mutant has 25% of wild type activity
R562Q
-
the mutant has 23% of wild type activity
R632K
-
the mutant enzyme has 72% of wild type activity
R632Q
-
the mutant enzyme has 58% of wild type activity
R655A
-
the mutant has 31% of wild type activity
R794A
-
the mutant has 124% of wild type activity
R808A
-
the mutant has 142% of wild type activity
S186A/D187A
-
modest defect, grows well at 19, 30 and 34C, but very slowly at 37C
S569A
-
the mutant enzyme has 39% of wild type activity
V750A
-
the mutant enzyme has 59% of wild type activity
W189A/L190A
-
temperature-sensitive vD12 allele, does not grow at 37C
W677A
-
the mutant enzyme has 4% of wild type activity
Y258A-V259A
-
modest defect, grows well at 19, 30 and 34C, but slowly at 37C
Y39A/G40A
-
temperature-sensitive vD12 allele, does not grow at 37C
Y555A
-
the mutant has 2% of wild type activity
Y555F
-
the mutant has 94% of wild type activity
Y555L
-
the mutant has 2% of wild type activity
Y555S
-
the mutant has 4% of wild type activity
Y608A
-
the mutant enzyme has 3% of wild type activity
Y608F
-
the mutant enzyme has 74% of wild type activity
Y608L
-
the mutant enzyme has 62% of wild type activity
Y608S
-
the mutant enzyme has 2% of wild type activity
Y683A
-
in D1 fragment, amino acids 498-844, 0.05% activity remaining, unable to crosslink the cap in presence of S-adenosyl-L-homocysteine
Y683F
-
in D1 fragment, amino acids 498-844, 4% activity remaining
Y683S
-
in D1 fragment, amino acids 498-844, catalytically defective, unable to crosslink the cap in presence of S-adenosyl-L-homocysteine
Y764A
-
the mutant enzyme has 18% of wild type activity
Y836A
-
the mutant enzyme has 15% of wild type activity
E1764A
-
mutation abolishes both guanine-N-7 (G-N-7) and ribose 2'-O methylation methylation
F1691A
-
mutation abolishes both guanine-N-7 (G-N-7) and ribose 2'-O methylation. Alanine substitution results in recombinant virus that does not form plaques and significant diminished viral replication
F1691W
-
mutation does not significantly affect the size of viral plaques. Mutant replicates as wild-type VSV. Mutant exhibits 82% of wild-type guanine-N-7 (G-N-7) methylation
F1691Y
-
mutation does not significantly affect the size of viral plaques. Mutant replicates as wild-type VSV. Mutant exhibits 85% of wild-type guanine-N-7 (G-N-7) methylation
F1745A
-
mutant shows a moderate defect in guanine-N-7 (G-N-7) methylation
F1816A
-
mutant does not show a significant defect in guanine-N-7 (G-N-7) methylation
L1757A
-
mutant does not show a significant defect in guanine-N-7 (G-N-7) methylation
N1692A
-
mutant shows a moderate defect in guanine-N-7 (G-N-7) methylation
P1709A
-
mutant does not show a significant defect in guanine-N-7 (G-N-7) methylation
S1693A
-
mutant shows a moderate defect in guanine-N-7 (G-N-7) methylation
S1827A
-
mutant shows a moderate defect in guanine-N-7 (G-N-7) methylation
W1744A
-
mutant does not show a significant defect in guanine-N-7 (G-N-7) methylation
Y1650A
-
mutation abolishes both guanine-N-7 (G-N-7) and ribose 2'-O methylation. Alanine substitution results in recombinant virus that does not form plaques and significant diminished viral replication
Y1650F
-
mutation does not significantly affect the size of viral plaques. Mutant replicates as wild-type VSV. Mutant exhibits 88% of wild-type guanine-N-7 (G-N-7) methylation
Y1650W
-
mutation does not significantly affect the size of viral plaques. Mutant replicates as wild-type VSV. Mutant exhibits 90% of wild-type guanine-N-7 (G-N-7) methylation
Y1835A
-
mutant does not show a significant defect in guanine-N-7 (G-N-7) methylation
E149A
-
the mutant shows about 1% N7 MTase activity as compared to the wild type protein
E34A
-
the mutant shows about 70% N7 MTase activity as compared to the wild type protein
F133A
-
the mutant shows about 99% N7 MTase activity as compared to the wild type protein
H42A
-
the mutant shows wild type N7 MTase activity
K21A
-
the mutant shows about 95% N7 MTase activity as compared to the wild type protein
K41A
-
the mutant shows about 90% N7 MTase activity as compared to the wild type protein
K45A
-
the mutant shows about 105% N7 MTase activity as compared to the wild type protein
L16A
-
the mutant shows about 110% N7 MTase activity as compared to the wild type protein
L184A
-
the mutant shows about 85% N7 MTase activity as compared to the wild type protein
Q114A
-
the mutant shows about 97% N7 MTase activity as compared to the wild type protein
R213A
-
the mutant shows about 30% N7 MTase activity as compared to the wild type protein
R37A
-
the mutant shows about 5% N7 MTase activity as compared to the wild type protein
R44A
-
the mutant shows about 65% N7 MTase activity as compared to the wild type protein
R57A
-
the mutant shows about 10% N7 MTase activity as compared to the wild type protein
R84A
-
the mutant shows about 3% N7 MTase activity as compared to the wild type protein
T216A
-
the mutant shows wild type N7 MTase activity
V55A
-
the mutant shows about 50% N7 MTase activity as compared to the wild type protein
W87A
-
the mutant shows about 10% N7 MTase activity as compared to the wild type protein
Y220A
-
the mutant shows about 30% N7 MTase activity as compared to the wild type protein
Y254A
-
the mutant shows about 110% N7 MTase activity as compared to the wild type protein
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
drug development
-
cap methylation is a principal target of the antifungal activity of sinefungin
synthesis
combination of chemical synthesis and enzymatic methylation in order to produce large amounts of RNA oligonucleotides carrying a cap-0 or cap-1. Short RNAs are synthesized on solid support by the phosphoramidite 2'-O-pivaloyloxymethyl chemistry. The cap structure is then coupled by the addition of GDP after phosphorylation of the terminal 5'-OH and activation by imidazole. GpppN-RNAs or GpppN2'-Om-RNAs are purified before the N7-methyl group is added using the human (guanine-N7)-methyl transferase to yield 7mGpppN-RNAs (cap-0) or 7mGpppN29-Om-RNAs (cap-1)
Show AA Sequence (1102 entries)
Please use the Sequence Search for a specific query.