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Information on EC 5.4.99.20 - 23S rRNA pseudouridine2457 synthase and Organism(s) Escherichia coli and UniProt Accession P75966

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     5 Isomerases
         5.4 Intramolecular transferases
             5.4.99 Transferring other groups
                5.4.99.20 23S rRNA pseudouridine2457 synthase
IUBMB Comments
The enzyme modifies uridine2457 in a stem of 23S RNA in Escherichia coli.
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This record set is specific for:
Escherichia coli
UNIPROT: P75966
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The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
23S rRNA uridine2457
=
23S rRNA pseudouridine2457
Synonyms
pseudouridine synthase, RluE, YmfC, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
23S rRNA-uridine2457 uracil mutase
The enzyme modifies uridine2457 in a stem of 23S RNA in Escherichia coli.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
23S rRNA uridine2457
23S rRNA pseudouridine2457
show the reaction diagram
additional information
?
-
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
23S rRNA uridine2457
23S rRNA pseudouridine2457
show the reaction diagram
additional information
?
-
RluE acts as a stand-alone, highly specific enzyme forming the universally conserved pseudouridine at position 2457, located in helix 89 (H89) of the 23S rRNA in the peptidyltransferase center, detailed structure-function analysis to determine the structural elements both in RluE and in 23S rRNA required for RNA-protein interaction and pseudouridine formation
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-
?
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
dissociation constants (KD) of wild-type and mutant enzymes, kinetics, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.7
23S rRNA uridine2457
pH 7.5, 37°C
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
pseudouridylation is catalyzed by two different systems: stand-alone pseudouridine synthases found in all domains of life and H/ACAribonucleoproteins (RNPs) that are only present in archaea and eukaryotes. All pseudouridine synthases are classified into six families based on the presence of common motifs, which are named after representative enzymes, namely, RluA, RsuA, TruA, TruB, TruD and Pus10
malfunction
pseudouridine2457 is absent in a ymfC deletion strain, it reappears when the deletion strain is transformed by a rescue plasmid carrying the ymfC structural gene
physiological function
Pseudouridines are not only found in rRNA but in all types of cellular RNA including mRNAs, tRNAs, small nuclear RNAs, small nucleolar RNAs and long-noncoding RNAs. In general, pseudouridines are thought to play a role in RNA stabilization, structure and correct RNA folding. RluE is a faster pseudouridine synthase than other enzymes which likely enables it to act in the early stages of ribosome formation. Molecular mechanism of RluE forming a highly conserved pseudouridine during ribosome biogenesis, overview
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24900
24900, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
24900, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, crystal structures of the C-terminal, catalytic domain of Escherichia coli RluE at 1.2 A resolution and of full-length RluE at 1.6 A resolution. The crystals of the full-length enzyme contain two molecules in the asymmetric unit and in both molecules the N-terminal domain is disordered. The higher resolution, truncated RluE structure is used for analysis and for comparison to other pseudouridine synthases
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D79N
inactive mutant enzyme
D79T
inactive mutant enzyme
R153A/P155G/R158A/R160A
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
R196A
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
R31A/R32A/K33A
site-directed mutagenesis, the mutant shows pseudouridylation activity similar to wild-type
R60A/K61A
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
additional information
generation of the deletion mutants RluE DELTAN(1-35) lacking the N-terminal region deleted at residue 35 and RluE DELTAR158-K161 lacking loop L7-8. Mutant RluE DELTAN(1-35) shows pseudouridylation activity similar to wild-type, while the activity of mutant RluE DELTAR158-K161 is significantly reduced
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain AG1 (ME5305) by nickel affinity chromatography, ultrafiltration, and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene rluE, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain AG1 (ME5305)
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Pan, H.; Ho, J.D.; Stroud, R.M.; Finer-Moore, J.
The crystal structure of E. coli rRNA pseudouridine synthase RluE
J. Mol. Biol.
367
1459-1470
2007
Escherichia coli (P75966)
Manually annotated by BRENDA team
Del Campo, M.; Kaya, Y.; Ofengand, J.
Identification and site of action of the remaining four putative pseudouridine synthases in Escherichia coli
RNA
7
1603-1615
2001
Escherichia coli (P75966)
Manually annotated by BRENDA team
Tillault, A.S.; Schultz, S.K.; Wieden, H.J.; Kothe, U.
Molecular determinants for 23S rRNA recognition and modification by the E. coli pseudouridine synthase RluE
J. Mol. Biol.
430
1284-1294
2018
Escherichia coli (P75966)
Manually annotated by BRENDA team