Information on EC 5.4.99.B22 - multisite-specific tRNA pseudouridine synthase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
5.4.99.B22
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
multisite-specific tRNA pseudouridine synthase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
tRNA uridine26 = tRNA pseudouridine26
show the reaction diagram
tRNA uridine27 = tRNA pseudouridine27
show the reaction diagram
tRNA uridine28 = tRNA pseudouridine28
show the reaction diagram
tRNA uridine30 = tRNA pseudouridine30
show the reaction diagram
tRNA uridine34 = tRNA pseudouridine34
show the reaction diagram
tRNA uridine35 = tRNA pseudouridine35
show the reaction diagram
tRNA uridine36 = tRNA pseudouridine36
show the reaction diagram
tRNA uridine65 = tRNA pseudouridine65
show the reaction diagram
tRNA uridine67 = tRNA pseudouridine67
show the reaction diagram
SYSTEMATIC NAME
IUBMB Comments
multisite-specific tRNA-uridine uracil mutase
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
human pre-tRNASer(UGA) uridine27
human pre-tRNASer(UGA) pseudouridine27
show the reaction diagram
-
-
-
?
human tRNAMet uridine27
human tRNAMet pseudouridine27
show the reaction diagram
-
-
-
-
?
mouse tRNAIle(UAU) uridine30
mouse tRNAIle(UAU) pseudouridine30
show the reaction diagram
-
-
-
-
?
mouse tRNAMet uridine27
mouse tRNAMet pseudouridine27
show the reaction diagram
-
-
-
-
?
pre-tRNAIle(UAU) uridine27
pre-tRNAIle(UAU) pseudouridine27
show the reaction diagram
-
-
-
-
?
pre-tRNAIle(UAU) uridine30
pre-tRNAIle(UAU) pseudouridine30
show the reaction diagram
-
-
-
-
?
pre-tRNAIle(UAU) uridine34
pre-tRNAIle(UAU) pseudouridine34
show the reaction diagram
-
-
-
-
?
pre-tRNAIle(UAU) uridine36
pre-tRNAIle(UAU) pseudouridine36
show the reaction diagram
-
-
-
-
?
tRNAArg(ACG) uridine1
tRNAArg(ACG) pseudouridine1
show the reaction diagram
tRNAIle uridine27
tRNAIle pseudouridine27
show the reaction diagram
-
-
-
-
?
tRNAIle(UAU) uridine27
tRNAIle(UAU) pseudouridine27
show the reaction diagram
tRNAIle(UAU) uridine34
tRNAIle(UAU) pseudouridine34
show the reaction diagram
tRNAIle(UAU) uridine36
tRNAIle(UAU) pseudouridine36
show the reaction diagram
tRNATrp(CCA) uridine26
tRNATrp(CCA) pseudouridine26
show the reaction diagram
tRNATrp(CCA) uridine27
tRNATrp(CCA) pseudouridine27
show the reaction diagram
tRNATrp(CCA) uridine28
tRNATrp(CCA) pseudouridine28
show the reaction diagram
tRNATrp(CCA) uridine34
tRNATrp(CCA) pseudouridine34
show the reaction diagram
-
-
-
?
tRNATrp(CCA) uridine35
tRNATrp(CCA) pseudouridine35
show the reaction diagram
-
-
-
?
tRNATrp(CCA) uridine36
tRNATrp(CCA) pseudouridine36
show the reaction diagram
-
-
-
?
tRNATrp(CCA) uridine65
tRNATrp(CCA) pseudouridine65
show the reaction diagram
-
-
-
?
tRNATrp(CCA) uridine67
tRNATrp(CCA) pseudouridine67
show the reaction diagram
-
-
-
?
tRNAVal uridine27
tRNAVal pseudouridine27
show the reaction diagram
-
-
-
-
?
yeast pre-tRNAIle uridine
yeast pre-tRNAIle pseudouridine
show the reaction diagram
-
possible modification sites are uridine 27, uridine30, uridine34 or uridine36
-
-
?
yeast tRNASer(UAU) uridine27
yeast tRNASer(UAU) pseudouridine27
show the reaction diagram
-
-
-
?
yeast tRNASer(UAU) uridine34
yeast tRNASer(UAU) pseudouridine34
show the reaction diagram
-
-
-
?
yeast tRNASer(UAU) uridine36
yeast tRNASer(UAU) pseudouridine36
show the reaction diagram
-
-
-
?
yeast tRNAVal(UAC) uridine27
yeast tRNAVal(UAC) pseudouridine27
show the reaction diagram
-
-
-
?
yeast tRNAVal(UAC) uridine32
yeast tRNAVal(UAC) pseudouridine32
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
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
-
the complex of Pus1 with tRNA is sensitive to high ionic strength. The optimal concentration of KCl for Pus1 binding is 100 mM. Above 400 mM KCl, no complex is detected
Zinc
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contains one atom of zinc per 63000 Da monomer. This zinc ion could not be removed by treatment with EDTA or urea. However, a zinc-depleted enzyme is obtained after prolonged dialysis against the specific chelating agent 1,10-phenanthroline. Removal of the zinc ion results in inactivation of the enzyme with concomitant loss of its ability to bind tRNA. Structural role for the zinc in the tRNA-pseudouridine synthetase Pus1
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
-
the complex Pus1/tRNAVal is sensitive to the presence of EDTA
KCl
-
the complex of Pus1 with tRNA is sensitive to high ionic strength. The optimal concentration of KCl for Pus1 binding is 100 mM. Above 400 mM KCl, no complex is detected
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00074
tRNAIle uridine27
-
pH and temperature not specified in the publication
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0.00042
tRNAVal uridine27
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pH and temperature not specified in the publication
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0.000032 - 0.000168
yeast pre-tRNAIle uridine
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0083
tRNAIle uridine27
Saccharomyces cerevisiae
-
pH and temperature not specified in the publication
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0.0067
tRNAVal uridine27
Saccharomyces cerevisiae
-
pH and temperature not specified in the publication
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.26
tRNAIle uridine27
Saccharomyces cerevisiae
-
pH and temperature not specified in the publication
41384
15.95
tRNAVal uridine27
Saccharomyces cerevisiae
-
pH and temperature not specified in the publication
41383
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
Pus1p is an intranuclear protein
Manually annotated by BRENDA team
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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study of the oligomerisation in solution of purified recombinant yeast tRNA:pseudouridine synthetase Pus1 in the presence or absence oftRNA. The presence of the neutral detergent dodecyl-beta-D-maltoside helps in avoiding the oligomerization/inactivation of the purified Pus1 in the absence of tRNA
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Saccharomyces cerevisiae
recombinant Pus1, tagged with six histidine residues at its N-terminus is expressed in Escherichia coli
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the PUS1 ORF is tagged at its amino-terminal end with six histidines and overexpressed in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R116A
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the mutant exhibits some activity with the yeast pre-tRNAIle substrate. Low activity with the mouse tRNAMet substrate
R116C
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the activity of the mutant enzyme with the yeast pre-tRNAIle substrate is approximately one-third that of the wild-type levels. No activity with the mouse tRNAMet substrate
R116E
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the mutant is inactive with with the yeast pre-tRNAIle substrate. No activity with the mouse tRNAMet substrate
R116G
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the mutant exhibits some activity with the yeast pre-tRNAIle substrate. No activity with the mouse tRNAMet substrate
R116H
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the mutant is inactive with with the yeast pre-tRNAIle substrate. Low activity with the mouse tRNAMet substrate
R116K
-
the activity of the mutant enzyme with the yeast pre-tRNAIle substrate is approximately one-third that of the wild-type levels
R116N
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the mutant exhibits some activity with the yeast pre-tRNAIle substrate. No activity with the mouse tRNAMet substrate
R116Q
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the mutant exhibits some activity with the yeast pre-tRNAIle substrate. No activity with the mouse tRNAMet substrate
R116S
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the mutant exhibits some activity with the yeast pre-tRNAIle substrate. No activity with the mouse tRNAMet substrate
R116W
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the mutant is inactive with with the yeast pre-tRNAIle substrate. Low activity with the mouse tRNAMet substrate
Y173C
-
mutant enzyme retains some activity with yeast pre-tRNAIle. The activity is relatively low
Y173F
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mutant enzyme is minimally activity with yeast pre-tRNAIle; mutant enzyme retains some activity with yeast pre-tRNAIle. The activity is relatively low
Y173G
-
mutant enzyme retains some activity with yeast pre-tRNAIle. The activity is relatively low
Y173S
-
mutant enzyme is minimally activity with yeast pre-tRNAIle
Y173T
-
mutant enzyme is minimally activity with yeast pre-tRNAIle; mutant enzyme retains some activity with yeast pre-tRNAIle. The activity is relatively low
D112A
-
inactive mutant enzyme