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Information on EC 5.4.99.28 - tRNA pseudouridine32 synthase
for references in articles please use BRENDA:EC5.4.99.28
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EC Tree 5 Isomerases
5.4 Intramolecular transferases
5.4.99 Transferring other groups
5.4.99.28 tRNA pseudouridine32 synthase
IUBMB Comments
The dual-specificity enzyme from Escherichia coli also catalyses the formation of pseudouridine746 in 23S rRNA . cf. EC 5.4.99.29 (23S rRNA pseudouridine746 synthase).
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
- 5.4.99.28
- synthases
- rrna
-
dual-specificity
-
pseudouridylating
- 5-fluorouridine
showSynonyms
pseudouridine synthase rlua, rib2/pus8p, trna:psi32-synthase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
tRNA uridine32 = tRNA pseudouridine32
RluA recognizes its substrates indirectly by probing RNA loops for their ability to adopt the reorganized fold
tRNA uridine32 = tRNA pseudouridine32
RluA does not require thiol(ate) groups to effect catalysis
tRNA uridine32 = tRNA pseudouridine32
-
-
-
-
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SYSTEMATIC NAME
IUBMB Comments
tRNA-uridine32 uracil mutase
The dual-specificity enzyme from Escherichia coli also catalyses the formation of pseudouridine746 in 23S rRNA [5]. cf. EC 5.4.99.29 (23S rRNA pseudouridine746 synthase).
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CAS REGISTRY NUMBER
COMMENTARY
430429-15-5
-
61506-89-6
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
tRNA uridine32
tRNA pseudouridine32
Substrates: RluA is the only synthase for formation of pseudouridine746 in 23S tRNA
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: stem-loop RNAs are good substrates for RluA
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: -
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: substrate: Escherichia coli tRNAPhe
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: substrate tRNAPhe. The enzyme is also responsible for the formation of pseudouridine746 in 23S rRNA
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: Rib2/Pus8p acts specifically on cytoplasmic tRNAs, whereas Pus9p modifies the same position mostly in mitochondrial tRNAs
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: -
Products: -
Products: -
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
tRNA uridine32
tRNA pseudouridine32
Substrates: RluA is the only synthase for formation of pseudouridine746 in 23S tRNA
Products: -
Products: -
?
tRNA uridine32
tRNA pseudouridine32
Substrates: Rib2/Pus8p acts specifically on cytoplasmic tRNAs, whereas Pus9p modifies the same position mostly in mitochondrial tRNAs
Products: -
Products: -
?
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000108
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, wild-type enzyme
-
0.000108
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme
-
0.000157
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A/C128A
-
0.00016
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C128A
-
0.000308
tRNA uridine32
pH 7.5, 37°C, anticodon stem-loop of Escherichia coli tRNAPhe (17-mer)
-
0.000318
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P29G
-
0.00042
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A
-
0.000455
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P29L
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.051
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P29L
-
0.061
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P29G
-
0.068
tRNA uridine32
pH 7.5, 37°C, anticodon stem-loop of Escherichia coli tRNAPhe (17-mer)
-
0.07
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A
-
0.074
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A/C128A
-
0.099
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, wild-type enzyme
-
0.099
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme
-
0.15
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C128A
-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
112.1
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P29L
-
166.7
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A
-
191.8
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P29G
-
220
tRNA uridine32
pH 7.5, 37°C, anticodon stem-loop of Escherichia coli tRNAPhe (17-mer)
-
471.3
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A/C128A
-
916.7
tRNA uridine32
pH 7.5, 37°C, Escherichia coli tRNAPhe, wild-type enzyme
-
916.7
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme
-
937.5
tRNA uridine32
pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C128A
-
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
malfunction
the rluA deletion mutant fails to form either 23S RNA pseudouridine746 or tRNA pseudouridine32. Replacement of rluA in trans on a rescue plasmid restores both pseudouridines. There is no difference in exponential growth rate between wild-type and MG1655(rluA-) either in rich or minimal medium at 24, 37, or 42°C, but when both strains are grown together, a strong selection against the deletion strain is observed
malfunction
Rib2/Pus8p acts on cytoplasmic tRNAs, whereas Pus9p modifies the same position in mitochondrial tRNAs. The DELTArib2/pus8/DELTApus9, is viable only in the presence of 15 mg/liter riboflavin in the growth medium
Show AA Sequence and Transmembrane Helices (2182 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
vapor diffusion method. 2.05 A resolution structure of RluA bound to a substrate RNA comprising the anticodon stem loop of tRNAPhe reveals that enzyme binding induces a dramatic reorganization of the RNA
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C117A
kcat is 1.4fold lower than wild-type value, KM is 3.9fold higher than wild-type value
C117A/A128A
kcat is 1.3fold lower than wild-type value, KM is 1.5fold higher than wild-type value
C128A
kcat is 1.5fold higher than wild-type value, KM is 1.5fold higher than wild-type value
P29G
kcat-value is 1.6fold lower than wild-type value. KM-value is 2.9fold higher than wild-type value
P29L
kcat-value is 1.9fold lower than wild-type value. KM-value is 4.2fold higher than wild-type value
D112A
mutant does not pseudouridine32 in formation in cytoplasmic tRNAGly of the DELTArib2/pus8/DELTApus9 strain
additional information
additional information
substitution of cysteine for amino acids with nonnucleophilic side chains does not significantly alter the catalytic activity
additional information
Pus9P lacking the 23 N-terminal amino acids is inactive
additional information
Pus9P lacking the 23 N-terminal amino acids is inactive
additional information
-
Pus9P lacking the 23 N-terminal amino acids is inactive
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ramamurthy, V.; Swann, S.; Spedaliere, C.; Mueller, E.
Role of cysteine residues in pseudouridine synhases of different families
Biochemistry
38
13106-13111
1999
Escherichia coli (P0AA37)
brenda
Hamilton, C.S.; Greco, T.M.; Vizthum, C.A.; Ginter, J.M.; Johnston, M.V.; Mueller, E.G.
Mechanistic investigations of the pseudouridine synthase RluA using RNA containing 5-fluorouridine
Biochemistry
45
12029-12038
2006
Escherichia coli (P0AA37)
brenda
Hoang, C.; Chen, J.; Vizthum, C.A.; Kandel, J.M.; Hamilton, C.S.; Mueller, E.G.; Ferre-DAmare, A.R.
Crystal structure of pseudouridine synthase RluA: indirect sequence readout through protein-induced RNA structure
Mol. Cell
24
535-545
2006
Escherichia coli (P0AA37)
brenda
Spedaliere, C.J.; Hamilton, C.S.; Mueller, E.G.
Functional importance of motif I of pseudouridine synthases: mutagenesis of aligned lysine and proline residues
Biochemistry
39
9459-9465
2000
Escherichia coli (P0AA37)
brenda
Raychaudhuri, S.; Niu, L.; Conrad, J.; Lane, B.G.; Ofengand, J.
Functional effect of deletion and mutation of the Escherichia coli ribosomal RNA and tRNA pseudouridine synthase RluA
J. Biol. Chem.
274
18880-18886
1999
Escherichia coli (P0AA37), Escherichia coli
brenda
Wrzesinski, J.; Nurse, K.; Bakin, A.; Lane, B.G.; Ofengand, J.
A dual-specificity pseudouridine synthase: an Escherichia coli synthase purified and cloned on the basis of its specificity for psi 746 in 23S RNA is also specific for psi 32 in tRNA(phe)
RNA
1
437-448
1995
Escherichia coli (P0AA37)
brenda
Behm-Ansmant, I.; Grosjean, H.; Massenet, S.; Motorin, Y.; Branlant, C.
Pseudouridylation at position 32 of mitochondrial and cytoplasmic tRNAs requires two distinct enzymes in Saccharomyces cerevisiae
J. Biol. Chem.
279
52998-53006
2004
Saccharomyces cerevisiae (Q12362), Saccharomyces cerevisiae (Q12069), Saccharomyces cerevisiae
brenda
EXTERNAL LINKS (specific for EC-Number 5.4.99.28)
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