Information on EC 3.1.27.4 - ribonuclease U2

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.1.27.4
-
RECOMMENDED NAME
GeneOntology No.
ribonuclease U2
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Ap or Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
CAS REGISTRY NUMBER
COMMENTARY hide
37205-57-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
synthetic construct
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-
-
Manually annotated by BRENDA team
CBS 534.71
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
5'-(U)6-GA-(U)5-3' RNA + H2O
?
show the reaction diagram
synthetic construct
-
digestion of the platinated 5'-(U)6-GA-(U)5-3' RNA shows a reaction product corresponding to [UUUUUUGA + Pt(NH3)2 + H],+ depicting RNase cleavage 3' to an adenosine predicted to be involved in a cisplatin diadduct. The platinated G*A* RNA adduct is uniquely recognized and processed by RNase U2, whereas other purine-Pt(II) adducts are not. The basis for this selectivity may arise from the RNase's known A - G preference, allowing the endonuclease in some cases to partially recognize the 3' platinated A despite platinum modification
-
-
?
RNA + H2O
3'-Phosphomononucleotides + 3'-phosphooligonucleotides + 2',3'-cyclic nucleotide phosphates
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
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
region Asp29-Asp37 winds around a calcium ion
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adenosine
-
-
Ethoxyformic anhydride
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poly-L-lysine
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spermidine
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-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2 - 0.234
ApA
-
-
0.075
ApAp
-
-
0.077
ApCp
-
-
0.041
ApGp
-
-
0.062
ApUp
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-
0.2 - 0.23
RNA
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0852
adenosine 3'-(1-naphthyl phosphate)
Ustilago sphaerogena
-
-
0.00983
adenosine 3'-(benzyl phosphate)
Ustilago sphaerogena
-
-
0.00317
adenosine 3'-(methyl phosphate)
Ustilago sphaerogena
-
-
3.33
ApA
Ustilago sphaerogena
-
-
24.3
ApAp
Ustilago sphaerogena
-
-
23.8
ApC
Ustilago sphaerogena
-
-
96.2
ApCp
Ustilago sphaerogena
-
-
229
ApCpGp
Ustilago sphaerogena
-
-
6.33
ApG
Ustilago sphaerogena
-
-
37.2
ApGp
Ustilago sphaerogena
-
-
7.33
ApU
Ustilago sphaerogena
-
-
55
ApUp
Ustilago sphaerogena
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-
141 - 149
ApUpG
282
ApUpGp
Ustilago sphaerogena
-
-
59.5
CpApGp
Ustilago sphaerogena
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-
14.8
UpApGp
Ustilago sphaerogena
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-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
specific activity increases 78fold after purification from crude extracts
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 4.5
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adenosine 2',3'-cyclic phosphate + uridine
7 - 7.5
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RNA
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10000
-
Ustilago sphaerogena, amino acid analysis
12400
-
Ustilago sphaerogena, sedimentation equilibrium
12590
-
mutant C1S/C54S, MALDI-TOF
13030
-
wild type, MALDI-TOF
13330
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mutant H101Q, MALDI-TOF, posttranslational modification suggested
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 11000, SDS-PAGE of nonreduced protein, x * 22000, SDS-PAGE of reduced protein
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
heterotrimeric complex of (YefM)2-YoeB antitoxin-toxin pair and free YoeB structure. Conformational rearrangement of YoeB RNase catalytic site upon binding of YefM
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isoAsp containing ribonuclease U2B, hanging drop vapor diffusion method, 0.004 ml of 10 mg/ml protein in 50 mM sodium citrate, pH 4.5, containing 50 mM sodium chloride, are mixed with 0.004 ml of reservoir solution containing 0.84 M sodium dihydrogen phosphate and 1.26 M dipotassium hydrogen phosphate, pH 6.9, equilibration against 0.4 ml reservoir solution, 20°C, 2 days, X-ray diffraction structure determination and analysis at 1.32 A resolution
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ribonuclease U2 complexed with adenosine 3'-monophosphate, hanging drop vapour diffusion method, 0.005 ml of protein solution containing 20 mg/ml RNase U2A and 8 mM adenosine 3'-monophosphate, pH 4.5, is mixed with 0.005 ml of reservoir solution containing 12.5% w/w PEG 8000, 200 mM calcium acetate, 100 mM sodium cacodylate, pH 3.75, 12.5% w/w PEG 8000, equilibration against 0.4 ml reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 0.96-0.99 A resolution
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RNase U2A in complex with 2'-adenylic acid and Ca2+ ions, X-ray diffraction structure determination and analysis at 1.03 A resolution
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
-
unstable above
100
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unstable above
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
3 Months, 0-4°C
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native RNase U2A by anion-exchange and 5'-adenylate aminohexyl affinity chromatography
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purification of recombinant enzyme using several chromatographic steps, including affinity chromatography on 2´-5'-ADP-Sepharose
purification of recombinant enzyme using several chromatographic steps, including DEAE-cellulose and affinity chromatography on 2'-5'-ADP-Sepharose
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Pichia pastoris, different signal-peptide cleavage processing sites for different recombinant proteins
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expression in Escherichia coli and Pichia pastoris
expression in Pichia pastoris
Pac1 endonuclease cleavage is required for U3 snoRNA gene transcription termination and RNA maturation, the degree of cleavage correlates closely with both RNA maturation and transcript termination. U3B snoRNA gene expression system containing a tagged Schizosaccharomyces pombe snU32 locus expressed under its own promoter and the dominant plasmid-derived U3B snoRNA
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H83Q
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no enzymic activity
R65A
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no enzymic activity
Y84A
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no enzymic activity
Y84F
-
reduced enzymic activity
C1S/C54S
-
lacking one disulfide bridge, 60% of wild-type activity
H101Q
-
devoid of detectable activity