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Information on EC 2.1.1.199 - 16S rRNA (cytosine1402-N4)-methyltransferase and Organism(s) Escherichia coli and UniProt Accession P60390

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EC Tree
IUBMB Comments
RsmH catalyses the N4-methylation of cytosine1402 and RsmI (EC 2.1.1.198) catalyses the 2'-O-methylation of cytosine1402 in 16S rRNA. Both methylations are necessary for efficient translation initiation at the UUG and GUG codons.
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This record set is specific for:
Escherichia coli
UNIPROT: P60390
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The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Reaction Schemes
hide
S-adenosyl-L-methionine
+
cytosine1402 in 16S rRNA
=
S-adenosyl-L-homocysteine
+
N4-methylcytosine1402 in 16S rRNA
+
cytosine1402 in 16S rRNA
=
+
N4-methylcytosine1402 in 16S rRNA
Synonyms
methyltransferase RsmH, mraW, RsmH, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
methyltransferase RsmH
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:16S rRNA (cytosine1402-N4)-methyltransferase
RsmH catalyses the N4-methylation of cytosine1402 and RsmI (EC 2.1.1.198) catalyses the 2'-O-methylation of cytosine1402 in 16S rRNA. Both methylations are necessary for efficient translation initiation at the UUG and GUG codons.
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 + cytidine1402 in 16S rRNA
S-adenosyl-L-homocysteine + N4-methylcytidine1402 in 16S rRNA
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + cytosine1402 in 16S rRNA
S-adenosyl-L-homocysteine + N4-methylcytosine1402 in 16S rRNA
show the reaction diagram
the enzyme recognizes the 30S subunit as a substrate, but not the naked 16S rRNA or the tightly-coupled 70S ribosome
-
-
?
S-adenosyl-L-methionine + cytosine1402 in 16S rRNA
S-adenosyl-L-homocysteine + N4-methylcytosine1402 in 16S rRNA
show the reaction diagram
-
-
-
-
?
NATURAL SUBSTRATE
NATURAL 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 + cytidine1402 in 16S rRNA
S-adenosyl-L-homocysteine + N4-methylcytidine1402 in 16S rRNA
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + cytosine1402 in 16S rRNA
S-adenosyl-L-homocysteine + N4-methylcytosine1402 in 16S rRNA
show the reaction diagram
the enzyme recognizes the 30S subunit as a substrate, but not the naked 16S rRNA or the tightly-coupled 70S ribosome
-
-
?
S-adenosyl-L-methionine + cytosine1402 in 16S rRNA
S-adenosyl-L-homocysteine + N4-methylcytosine1402 in 16S rRNA
show the reaction diagram
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
the cofactor binds in the active site, structure, overview
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
strain DELTAmraW produces monomethylated cytosine1402 instead of dimethylated C1402 produced by the wild-type enzyme
physiological function
both the 2'-O-methylation (catalyzed by RsmI) and N4-methylation of C1402 (catalyzed by RsmH) are needed for efficient initiation at the UUG and GUG codon. m4Cm1402 in the 16S rRNA modulates the accuracy of P-site function. The modification is needed for efficient translation initiation at the UUG and GUG codons
malfunction
-
enzyme deletion affects efficiency of non-AUG initiation and the fidelity of translation
additional information
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
active enzyme in solution, structure analysis, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant His6-tagged RsmH in complex with S-adenosyl-L-methionine and cytidine in a 1:5:5 ratio, sitting drop vapour diffusion method, from 20 mM Tris, 100 mM NaCl, 5% v/v glycerol, 2 mM DTT, pH 8.0, room temperature, the best crystal is obtained in the mixture solution 1.0 M ammonium sulfate, 0.1 M HEPES, pH 6.5, and 0.5% w/v PEG 8, 000 in 3-4 days, X-ray diffraction structure determination and analysis at 2.25 A resolution
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged RsmH from Escherichia coli strain BL21 DE3 star by nickel affinity chromatigraphy and gel filtration
recombinant protein with N-terminal 6His-tags
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene rsmH, expression of His6-tagged RsmH in Escherichia coli strain BL21 DE3 star
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kimura, S.; Suzuki, T.
Fine-tuning of the ribosomal decoding center by conserved methyl-modifications in the Escherichia coli 16S rRNA
Nucleic Acids Res.
38
1341-1352
2010
Escherichia coli (P60390)
Manually annotated by BRENDA team
Wei, Y.; Zhang, H.; Gao, Z.Q.; Wang, W.J.; Shtykova, E.V.; Xu, J.H.; Liu, Q.S.; Dong, Y.H.
Crystal and solution structures of methyltransferase RsmH provide basis for methylation of C1402 in 16S rRNA
J. Struct. Biol.
179
29-40
2012
Escherichia coli (P60390), Escherichia coli
Manually annotated by BRENDA team
Zhao, M.; Wang, L.; Zhang, H.; Dong, Y.; Gong, Y.; Zhang, L.; Wang, J.
Purification, crystallization and preliminary crystallographic analysis of the 16S rRNA methyltransferase RsmI from Escherichia coli
Acta Crystallogr. Sect. F
70
1256-1259
2014
Escherichia coli
Manually annotated by BRENDA team
Lioy, V.S.; Goussard, S.; Guerineau, V.; Yoon, E.J.; Courvalin, P.; Galimand, M.; Grillot-Courvalin, C.
Aminoglycoside resistance 16S rRNA methyltransferases block endogenous methylation, affect translation efficiency and fitness of the host
RNA
20
382-391
2014
Escherichia coli, Escherichia coli BW25113
Manually annotated by BRENDA team