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Information on EC 4.6.1.19 - ribonuclease T2 and Organism(s) Homo sapiens and UniProt Accession O00584

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IUBMB Comments
A widely distributed family of related enzymes found in protozoans, plants, bacteria, animals and viruses that cleave ssRNA 3'-phosphate group with little base specificity. The enzyme catalyses a two-stage endonucleolytic cleavage. The first reaction produces 5'-hydroxy-phosphooligonucletides and 3'-phosphooligonucleotides ending with a 2',3'-cyclic phosphodiester, which are released from the enzyme. The enzyme then hydrolyses the cyclic products in a second reaction that takes place only when all the susceptible 3',5'-phosphodiester bonds have been cyclised. The second reaction is a reversal of the first reaction using the hydroxyl group of water instead of the 5'-hydroxyl group of ribose. The overall process is that of a phosphorus-oxygen lyase followed by hydrolysis to form the 3'-nucleotides.
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Homo sapiens
UNIPROT: O00584
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Word Map
The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
hide(Overall reactions are displayed. Show all >>)
+
=
an [RNA fragment]-3'-nucleoside-3'-phosphate
+
a 5'-hydroxy-ribonucleotide-3'-[RNA fragment]
Synonyms
rnase l, omega-1, rnase t2, rnaset2, rnase i, acid rnase, ribonuclease l, rnase rh, acid ribonuclease, rnase m, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acid ribonuclease
-
-
-
-
acid RNase
base-non-specific ribonuclease
-
-
-
-
Escherichia coli ribonuclease I
-
-
-
-
leukocytic-type acid RNase
-
-
non-base specific ribonuclease
-
-
-
-
nonbase-specific RNase
-
-
-
-
nonspecific RNase
-
-
-
-
nuclease, Escherichia coli ribo-, II
-
-
-
-
nuclease, ribo- (non-base specific)
-
-
-
-
ribonuclease (non-base specific)
-
-
-
-
ribonuclease II
-
-
-
-
ribonuclease M
-
-
-
-
ribonuclease N2
-
-
-
-
ribonuclease PP2
-
-
-
-
ribonuclease PP3
-
-
-
-
ribonuclease U4
-
-
-
-
ribonucleate 3'-oligonucleotide hydrolase
-
-
-
-
ribonucleate nucleotido-2'-transferase (cyclizing)
-
-
-
-
RNAase CL
-
-
-
-
RNase (non-base specific)
-
-
-
-
RNase II
-
-
-
-
RNase L
RNase LE
-
-
-
-
RNase LX
-
-
-
-
RNase M
-
-
-
-
RNase MC
-
-
-
-
RNase Ms
-
-
-
-
RNase T2
RNaseT2
S1-RNase
-
-
-
-
S2-RNase
-
-
-
-
S3-RNase
-
-
-
-
S4-RNase
-
-
-
-
S5-RNase
-
-
-
-
S6-RNase
-
-
-
-
S7-RNase
-
-
-
-
T2 ribonuclease
-
-
T2 RNase
-
-
additional information
-
the enzyme belongs to the RNaseT2 enzyme family
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
RNA + H2O = an [RNA fragment]-3'-nucleoside-3'-phosphate + a 5'-hydroxy-ribonucleotide-3'-[RNA fragment]
show the reaction diagram
mechanism of RNA recognition, binding and cleavage by RNase L, overview
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
[RNA] 5'-hydroxy-ribonucleotide-3'-[RNA fragment]-lyase (cyclicizing; [RNA fragment]-3'- nucleoside-2',3'-cyclophosphate-forming and hydrolysing)
A widely distributed family of related enzymes found in protozoans, plants, bacteria, animals and viruses that cleave ssRNA 3'-phosphate group with little base specificity. The enzyme catalyses a two-stage endonucleolytic cleavage. The first reaction produces 5'-hydroxy-phosphooligonucletides and 3'-phosphooligonucleotides ending with a 2',3'-cyclic phosphodiester, which are released from the enzyme. The enzyme then hydrolyses the cyclic products in a second reaction that takes place only when all the susceptible 3',5'-phosphodiester bonds have been cyclised. The second reaction is a reversal of the first reaction using the hydroxyl group of water instead of the 5'-hydroxyl group of ribose. The overall process is that of a phosphorus-oxygen lyase followed by hydrolysis to form the 3'-nucleotides.
CAS REGISTRY NUMBER
COMMENTARY hide
37278-25-4
-
9075-06-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
RNA + H2O
?
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
RNA + H2O
?
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
complete inhibition at 50 mM
Zn2+
complete inhibition at 50 mM
poliovirus RNA
-
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2',5'-linked oligoadenylates
two different binding sites in the enzyme's ANK domain and a third site in the N lobe of the KH domain, sensing mechanism, structure-function analysis, overview. The KH/KH and kinase extension nuclease (KEN)/KEN interfaces are important in 2',5'-oligoadenylate-dependent enzyme activation
-
2',5'-Oligoadenylate
2'-5'-linked oligoadenylates
-
natural 2'-5'-linked tetraadenylates and three tetramers with 3'-end AMP units replaced with ribo-, arabino- and xylo-configured phosphonate analogues of AMP are enzyme activators with equal potency. The enzyme is activated by the binding of 2'-5'-linked oligoadenylates to the N-terminal ankyrin repeat domain, which causes the inactive monomer to form a catalytically active homodimer
-
2'-5'-oligoadenylate
-
short polyadenylate molecules that are connected by unique 2'-5' linkages, collectively referred to as 2-5A. 2-5A induces dimerization via the ANK domain and this dimerization correlates with activation of ribonuclease activity
viral dsRNA
-
viral dsRNA provokes the synthesis of 2'-5' oligoadenylate, 2-5A, binding of 2-5A to ankyrin repeats of monomeric RNase L leads to RNase L dimerization and activation
-
additional information
-
a dephosphorylated form of 2',5'-oligoadenylate does not activate RNase L
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
enzyme levels at different kidney glomerular filtration rates of blood
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.6 - 6
assay at
6.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
assay at
22
-
assay at room temperature
37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 30
-
RNA cleavage assay
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10
-
above
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
neuroendocrine neoplasms of the lung, well differentiated and poorly differentiated
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
RNase T2 is located within a region of the long arm of chromosome six that undergoes deletion or rearrangement in many cancers, including some cases of adult T-cell leukemia, ATL
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans, its structure shows a typical fold for members of the T2 RNase family with seven alpha-helices and eight beta-strands constituting an alpha+beta motif, as most of the central anti-parallel beta-sheet is clearly separated in the sequence from the helical parts. The alpha + beta core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences
malfunction
mutations in the gene of human enzyme RNase T2 are associated with white matter disease of the human brain showing bilateral temporal lobe cysts and multifocal white matter lesions which lead to psychomotor impairments,spasticity and epilepsy
metabolism
there is a possible link between the hypoxic state experienced by cancer cells and the expression level of RNASET2. A role of this gene as a stress-response factor is postulated
evolution
-
involvement of the pseudokinase domain of the enzyme cofactor sensing, nucleotide binding, dimerization, and ribonuclease functions highlights the evolutionary adaptability of the eukaryotic protein kinase fold
malfunction
metabolism
the enzyme degrades mitochondrial RNAs and it is also responsible for selective degradation of the cytosolic rRNAs on the outer membrane. The degradation activity also has a positive effect on nuclear transcription of rRNAs, suggesting a compensatory feedback mechanism, and affects protein translations in and out of mitochondria
physiological function
additional information
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
RNT2_HUMAN
256
0
29481
Swiss-Prot
Secretory Pathway (Reliability: 2), Mitochondrion (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35000
-
recombinant truncated enzyme RNase L(1-306), gel filtration
37000
-
recombinant truncated enzyme RNase L(1-333), gel filtration
62100
-
recombinant truncated enzyme RNase L(1-333) with bound 2'-5'-oligoadenylate, gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 15000-16000
dimer
homodimer
-
active enzyme form
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
N-linked glycoprotein, enzyme activity is independent of glycosylation, deglycosylation by EndoH leaving one N-acetyl glucosamine residue at each of the N-glycosylation sites
glycoprotein
-
-
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified enzyme, hanging drop vapour diffusion method, mixing of 0.002 ml of 9.2 mg/ml protein in 20 mM acetate, pH 6.0, and 20 mM NaCl, with 0.002 ml of reservoir solution containing 0.2 M NH4NO3 and 15% PEG 3350, equilibration over 0.6 ml of reservoir solution, X-ray diffraction structure determination and analysis at 1.6 A resolution
analysis of two crystal structures of human enzyme
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H672N
K392R
-
the mutation in the kinase ATP binding site of the nezyme results in greatly impaired RNase activity
R163A
action potential aI clamp enzyme mutant, which shows reduced activity compared to the wild-type enzyme
R412A
KH/KH interface enzyme mutant, which shows reduced activity compared to the wild-type enzyme
R427A
activator recognition enzyme mutant, which shows reduced activity compared to the wild-type enzyme
R667A
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Mg2+ and ATP or ADP protect the enzyme from proteolytic cleavage
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme by metal affinity chromatography, deglycosylation, concanavalin A affinity chromatography and gel filtration
recombinant His6-tagged enzyme variants from Escherichia coli strain BL21(DE3) or Spodoptera frugiperda Sf9 insect cells by ultracentrifugation, nickel affinity chromatography and gel filtration, dialysis, anion exchange chromatography, clevage by TEV protease and again nickel affinity chromatography, gel filtration and
-
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and dialysis
-
using CL6B Blue Sepharose affinity resin, a MonoQ column
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant expression of His-tagged enzyme
expression of FLAG-tagged wild-type, R667A mutant, and NDELTA385 deletion enzyme mutant in HeLa-M cells
-
expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
-
gene RNASET2, real-time PCR expression analysis
-
recombinant expression of His6-tagged enzyme with or without TEV cleavage site inserted after residue 333 and the amino acid sequence of ZMP comprised residues 15 to 129 of zebrafish MDMX with two mutations L46V and V95L, in Escherichia coli strain BL21(DE3) or in Spodoptera frugiperda Sf9 insect cells. The zebrafish sequence ZMP is a unique dimerization inducing domain and represents a tool protein for solving crystal structures of inhibitors binding to the p53-interacting domain of MDMX
-
untagged recombinant human RNase L in a baculovirus vector is expressed in SF21 insect cells, into pcDNA3 vectors for stable transfection of HeLa cells
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
overexpression of RNASET2 in consequence of the activation of HIF-1alpha.
significantly higher expression of RNASET2, HIF-1alpha, and its target CA IX in poorly differentiated than in well-differentiated LuNENs
reduction in the level of RNase T2 by Tax protein from human T cell leukemia virus type 1
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
-
it will be important, for proving advantageous in vaccines, to determine whether mutations in the RNA structure associated with the inhibition of RNAse L attenuate polyviruses or other group C enteroviruses in vivo
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Naskalski, J.; Anyszek, T.; Surdacka, E.; Hoyer, B.; Sulowicz, W.
Acid ribonuclease and alkaline ribonuclease isoenzymes in plasma of patients with decreased glomerular filtration rate
Clin. Chem. Lab. Med.
42
279-282
2004
Homo sapiens
Manually annotated by BRENDA team
Han, J.Q.; Townsend, H.L.; Jha, B.K.; Paranjape, J.M.; Silverman, R.H.; Barton, D.J.
A phylogenetically conserved RNA structure in the poliovirus open reading frame inhibits the antiviral endoribonuclease RNase L
J. Virol.
81
5561-5572
2007
Homo sapiens
Manually annotated by BRENDA team
Luhtala, N.; Parker, R.
T2 Family ribonucleases: ancient enzymes with diverse roles
Trends Biochem. Sci.
35
253-259
2010
Aspergillus niger, Aspergillus oryzae, Bovine viral diarrhea virus 1, Classical swine fever virus, Entamoeba histolytica, Homo sapiens, Rhizopus niveus, Schistosoma mansoni
Manually annotated by BRENDA team
Polakowski, N.; Han, H.; Lemasson, I.
Direct inhibition of RNAse T2 expression by the HTLV-1 viral protein Tax
Viruses
3
1485-1500
2011
Homo sapiens
Manually annotated by BRENDA team
Kriz, M.; Snasel, J.; Kopecky, V.; Pav, O.; Rosenberg, I.; Stepanek, J.
Structural changes of human RNase L upon homodimerization investigated by Raman spectroscopy
Biochim. Biophys. Acta
1824
1039-1044
2012
Homo sapiens
Manually annotated by BRENDA team
Garvie, C.W.; Vasanthavada, K.; Xiang, Q.
Mechanistic insights into RNase L through use of an MDMX-derived multi-functional protein domain
Biochim. Biophys. Acta
1834
1562-1571
2013
Homo sapiens
Manually annotated by BRENDA team
de Leeuw, M.; Gonzalez, A.; Lanir, A.; Roiz, L.; Smirnoff, P.; Schwartz, B.; Shoseyov, O.; Almog, O.
The 1.8 A crystal structure of ACTIBIND suggests a mode of action for T2 ribonucleases as antitumorigenic agents
J. Med. Chem.
55
1013-1020
2012
Homo sapiens, Rattus norvegicus
Manually annotated by BRENDA team
Chakrabarti, A.; Ghosh, P.K.; Banerjee, S.; Gaughan, C.; Silverman, R.H.
RNase L triggers autophagy in response to viral infections
J. Virol.
86
11311-11321
2012
Homo sapiens
Manually annotated by BRENDA team
Huang, H.; Zeqiraj, E.; Dong, B.; Jha, B.K.; Duffy, N.M.; Orlicky, S.; Thevakumaran, N.; Talukdar, M.; Pillon, M.C.; Ceccarelli, D.F.; Wan, L.C.; Juang, Y.C.; Mao, D.Y.; Gaughan, C.; Brinton, M.A.; Perelygin, A.A.; Kourinov, I.; Guarne, A.; Silverman, R.H.; Sicheri, F.
Dimeric structure of pseudokinase RNase L bound to 2-5A reveals a basis for interferon-induced antiviral activity
Mol. Cell
53
221-234
2014
Homo sapiens, Sus scrofa
Manually annotated by BRENDA team
Thorn, A.; Steinfeld, R.; Ziegenbein, M.; Grapp, M.; Hsiao, H.H.; Urlaub, H.; Sheldrick, G.M.; Gaertner, J.; Kraetzner, R.
Structure and activity of the only human RNase T2
Nucleic Acids Res.
40
8733-8742
2012
Homo sapiens (O00584), Homo sapiens
Manually annotated by BRENDA team
Zhang, A.; Dong, B.; Doucet, A.J.; Moldovan, J.B.; Moran, J.V.; Silverman, R.H.
RNase L restricts the mobility of engineered retrotransposons in cultured human cells
Nucleic Acids Res.
42
3803-3820
2014
Homo sapiens
Manually annotated by BRENDA team
Han, Y.; Donovan, J.; Rath, S.; Whitney, G.; Chitrakar, A.; Korennykh, A.
Structure of human RNase L reveals the basis for regulated RNA decay in the IFN response
Science
343
1244-1248
2014
Homo sapiens (Q05823), Homo sapiens
Manually annotated by BRENDA team
Uccella, S.; La Rosa, S.; Scaldaferri, D.; Monti, L.; Maragliano, R.; Sorrenti, E.; Gariboldi, M.; Taramelli, R.; Sessa, F.; Acquati, F.
New insights into hypoxia-related mechanisms involved in different microvascular patterns of bronchopulmonary carcinoids and poorly differentiated neuroendocrine carcinomas. Role of ribonuclease T2 (RNASET2) and HIF-1alpha
Hum. Pathol.
79
66-76
2018
Homo sapiens (O00584)
Manually annotated by BRENDA team
Huang, J.; Liu, P.; Wang, G.
Regulation of mitochondrion-associated cytosolic ribosomes by mammalian mitochondrial ribonuclease T2 (RNASET2)
J. Biol. Chem.
293
19633-19644
2018
Homo sapiens (C0HKG5), Mus musculus (C0HKG5), Mus musculus
Manually annotated by BRENDA team