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Information on EC 3.1.26.4 - ribonuclease H and Organism(s) Homo sapiens and UniProt Accession O75792

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EC Tree
     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.26 Endoribonucleases producing 5'-phosphomonoesters
                3.1.26.4 ribonuclease H
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Select one or more organisms in this record: ?
This record set is specific for:
Homo sapiens
UNIPROT: O75792 not found.
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The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Endonucleolytic cleavage to a 5'-phosphomonoester
Synonyms
reverse transcriptase, ribonuclease h, rnase h2, rnase hii, rnaseh2a, rnaseh1, ribonuclease hi, ribonuclease h2, rnase hiii, ribonuclease h1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ribonuclease H2
-
endoribonuclease H
-
-
-
-
huRNaseH1
-
hybrid nuclease
-
-
-
-
hybrid ribonuclease
-
-
-
-
hybridase
-
-
-
-
hybridase (ribonuclease H)
-
-
-
-
nuclease, hybrid ribo-
-
-
-
-
nuclease, ribo-, H
-
-
-
-
P32
-
-
-
-
ribonuclease H
ribonuclease H1
-
-
ribonulease H1
-
RNA*DNA hybrid ribonucleotidohydrolase
-
-
-
-
RNase H
RNase H1
RNase H2
RNase HI
-
-
-
-
RNase HII
RNase HIII
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Endonucleolytic cleavage to a 5'-phosphomonoester
show the reaction diagram
substrate structure influence on enzyme activity, substrate binding, overview
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
9050-76-4
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D14R1D3:DNA18 + H2O
?
show the reaction diagram
-
-
-
?
RNA18:DNA18 + H2O
?
show the reaction diagram
-
-
-
?
DNA-2'-methoxyethoxy RNA hybrid
?
show the reaction diagram
-
chimeric substrates containing a central DNA region with flanking northern-biased 2'-methoxyethyl nucleotides hybridized to complementary RNA, enhanced cleavage rates are observed for the eastern-biased 2'-ara-fluorothymidine and bulge inducing N-methylthymidine modifications positioned at the 5'-DNA/3'-MOE junction as well as the southern-biased 2'-methylthiothymidine and conformationally flexible tetrafluoroindole modifications positioned at the 5'-MOE/3'-DNA junction, overview
-
-
?
DNA-2'-methoxyethoxy-antisense RNA hybrid + H2O
?
show the reaction diagram
-
2'-methoxyethoxy nucleotides, positioned at the 3' and 5' poles, into the antisense oligodeoxyribonucleotide of the heteroduplex to alter the helical geometry of the substrate
-
-
?
DNA-RNA duplex + H2O
?
show the reaction diagram
DNA-RNA hybrid + H2O
?
show the reaction diagram
-
-
-
-
?
DNA-RNA hybrid + H2O
DNA + 5'-phosphonucleotides
show the reaction diagram
DNA/RNA hybrid + H2O
?
show the reaction diagram
-
-
-
-
?
DNA12-RNA1-DNA27/DNA40 hybrid + H2O
?
show the reaction diagram
-
enzyme cleaves RNA20/DNA20 hybrid and DNA12-RNA1-DNA27/DNA40 hybrid substrates with similar efficiency
-
?
dsDNA oligonucleotide with a single ribose + H2O
dsDNA oligonucleotide with 1 nt gap + 5'-monophosphate ribonucleotide
show the reaction diagram
-
enzyme excises misincorporated ribonucleotides in DNA, enzyme places the first 5' nick, while the second 3' cut is made by FEN-1 protein
-
-
?
peptide nucleic acid - 2'-deoxy 2'-fluoroarabinonucleic acid hybrid + H2O
?
show the reaction diagram
-
chimeric oligomers possessing a single central peptide nucleic acid insert are capable of forming hybrid duplexes with complementary RNA, although with diminished thermal stability in comparison to the unmodified oligomers
-
-
?
peptide nucleic acid - DNA + H2O
?
show the reaction diagram
-
chimeric oligomers possessing a single central peptide nucleic acid insert are capable of forming hybrid duplexes with complementary RNA, although with diminished thermal stability in comparison to the unmodified oligomers
-
-
?
poly(rAdT) + H2O
?
show the reaction diagram
-
-
-
-
?
poly-rA/poly-dT + H2O
?
show the reaction diagram
-
products are short oligonucleotides with very few intermediate-sized oligonucleotides
-
?
RNA*2'F-ANA-DNA hybrid + H2O
?
show the reaction diagram
-
cleaves the RNA portion of hybrid duplexes of butyl-modified 2'F-ANA-DNA oligonucleotides containing acyclic interresidue units with complementary RNA
-
-
?
RNA*antisense-DNA hybrid + H2O
?
show the reaction diagram
-
cleaves the RNA portion of hybrid duplexes of modified antisense DNA oligonucleotides containing acyclic interresidue units with complementary RNA
-
-
?
RNA*DNA hybrid + H2O
?
show the reaction diagram
RNA-DNA heteroduplex + H2O
ribonucleotide 5'-phosphomonoester + ?
show the reaction diagram
-
-
-
-
?
RNA-DNA hybrid + H2O
?
show the reaction diagram
RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
show the reaction diagram
RNA20/DNA20 hybrid + H2O
?
show the reaction diagram
-
enzyme cleaves RNA20/DNA20 hybrid and DNA12-RNA1-DNA27/DNA40 hybrid substrates with similar efficiency
-
?
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
DNA-RNA hybrid + H2O
DNA + 5'-phosphonucleotides
show the reaction diagram
RNA*DNA hybrid + H2O
?
show the reaction diagram
-
cleaves the RNA portion
-
-
?
RNA-DNA heteroduplex + H2O
ribonucleotide 5'-phosphomonoester + ?
show the reaction diagram
-
-
-
-
?
RNA-DNA hybrid + H2O
?
show the reaction diagram
RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ni2+
-
exhibits only modest activity as cofactor
Zn2+
-
exhibits only modest activity as cofactor
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1R,3R,4R,5R,8S)-8-benzyloxy-1-benzyloxymethyl-5-benzyloxyamino-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
(1R,3R,4R,5R,8S)-8-benzyloxy-1-benzyloxymethyl-5-trifluoroacetamino-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
(1R,3R,4R,5S,8S)-8-benzyloxy-1-benzyloxymethyl-5-((methylthio)thiocarbonyl)oxy-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
(1R,3R,4R,5S,8S)-8-benzyloxy-1-benzyloxymethyl-5-(4-methylbenzoyl)-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
(1R,3R,4R,5S,8S)-8-benzyloxy-1-benzyloxymethyl-5-hydroxy-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane (20a) and (1R,3R,4R,5R,8S)-8-benzyloxy-1-benzyloxymethyl-5-hydroxy-3-(thymin-1-yl)-2-oxabicyclo[3.2.1]octane
-
-
(1R,3R,4R,5S,8S)-8-benzyloxy-1-benzyloxymethyl-5-methoxalyloxy-5-methyl-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
(1R,3R,4R,8S)-8-benzyloxy-1-benzyloxymethyl-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
(1R,3R,4R,8S)-8-benzyloxy-1-benzyloxymethyl-5-one-3-(thymin-1-yl)-2-oxa-bicyclo[3.2.1]octane
-
-
1,6-dihydroxy-4-methyl-5-(N-phenoxyethanimidoyl)pyridin-2(1H)-one
-
1,6-dihydroxy-5-[N-[(2-methoxyphenyl)methoxy]ethanimidoyl]-4-methylpyridin-2(1H)-one
-
1,6-dihydroxy-5-[N-[(4-methoxyphenyl)methoxy]ethanimidoyl]-4-methylpyridin-2(1H)-one
-
1-(2-O-acetyl-3,5-O-benzyl-4-C-cyanoethyl-beta-D-ribofuranosyl)-thymine
-
-
1-(3,5-O-benzyl-2-O-phenoxythiocarbonyl-4-C-propionaldehyde-beta-D-ribofuranosyl)thymineO-benzyloxime
-
-
1-(3,5-O-benzyl-4-C-cyanoethyl-2-O-hydroxyl-beta-D-ribofuranosyl)-thymine
-
-
1-(3,5-O-benzyl-4-C-propionaldehyde-2-O-hydroxyl-beta-D-ribofuranosyl)thymine O-benzyl oxime
-
-
2-(2,3-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
-
-
2-(2,5-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
-
-
2-(4,6-dimethyl-3-oxo-1,2-benzothiazol-2(3H)-yl)-N-propylacetamide
-
-
2-(4-chlorophenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
-
-
2-(6-fluoro-3-oxo-1,2-benzothiazol-2(3H)-yl)-N-(4-methylphenyl)acetamide
-
-
2-phenyl-1,2-benzothiazol-3(2H)-one
-
-
2-[(cyclopentylcarbonyl)amino]-4-ethyl-5-methylthiophene-3-carboxamide
-
-
3,5-di-O-benzyl-4-C-cyanoethyl-1,2-O-isopropylidene-alpha-D-ribofuranose
-
-
5-nitrofuran-2-carboxylic acid [[4-(4-bromophenyl)-thiazol-2-yl]-(tetrahydrofuran-2-ylmethyl)-carbamoyl]-methyl ester
-
derivative of 5-nitrofuran-2-carboxylic acid carbamoyl methyl ester. 20-25 microM effectively inhibit HIV-1 replication
5-[N-(4-fluorophenoxy)ethanimidoyl]-1,6-dihydroxy-4-methylpyridin-2(1H)-one
-
5-[N-[(2-aminophenyl)methoxy]ethanimidoyl]-1,6-dihydroxy-4-methylpyridin-2(1H)-one
-
5-[N-[(2-fluorophenyl)methoxy]ethanimidoyl]-1,6-dihydroxy-4-methylpyridin-2(1H)-one
-
5-[N-[(4-fluorophenyl)methoxy]ethanimidoyl]-1,6-dihydroxy-4-methylpyridin-2(1H)-one
-
6-(naphthalen-2-yl)-3-(pyridin-3-yl)[1,2,4]triazolo[3,4-b][1,3,4]thiadiazole
-
-
6-fluoro-2-(2-methylphenyl)-1,2-benzothiazol-3(2H)-one
-
-
6-fluoro-2-(4-methylphenyl)-1,2-benzothiazol-3(2H)-one
-
-
alpha-thujaplicin
-
i.e. 2-hydroxy-3-(1-methylethyl)-2,4,6-cycloheptatrien-1-one
antisense oligodeoxynucleotides
-
directed against RNA polymerase II, replication protein A, and Ha-ras, determination of response in expression levels of the enzyme type 1 and 2, overview
-
beta-thujaplicin
-
i.e. 2-hydroxy-4-(1-methylethyl)-2,4,6-cycloheptatrien-1-one
beta-thujaplicinol
-
i.e. 2,7-dihydroxy-4-(1-methylethyl)-2,4,6-cycloheptatrien-1-one
Ca2+
-
calcium ions generally inactivate the enzyme and abolish catalysis
Co2+
-
in presence of Mg2+, inhibition
Cu2+
-
in presence of Mg2+, inhibition
ellipticine
-
ethyl 6-hydroxy-2-methoxy-5,7-dioxo-5,6,7,8-tetrahydro-1,6-naphthyridine-8-carboxylate
-
Fe2+
-
in presence of Mg2+, inhibition
gamma-thujaplicin
-
i.e. 2-hydroxy-5-(1-methylethyl)-2,4,6-cycloheptatrien-1-one
KCl
-
half-maximal inhibition at 150 mM
manicol
-
i.e. 1,2,3,4-tetrahydro-2,7-dihydroxy-9-methyl-2-(1-methylethyl)-6H-benzocyclohepten-6-one
methyl 7-benzamido-2-hydroxy-1,3-dioxo-1,2,3,4-tetrahydroisoquinoline-4-carboxylate
-
Mg2+
-
Mg2+ is inhibitory at concentrations above 10 mM
Mn2+
-
in presence of Mg2+, strong inhibition
mtSSB
repressive effect of mtSSB, mildly at 20 nM, strongly at 120 nM
-
N-(5-benzyl-1,3,4-thiadiazol-2-yl)-2-ethylhexanamide
-
-
N-benzyl-2-(6-fluoro-3-oxo-1,2-benzothiazol-2(3H)-yl)acetamide
-
-
N-cyclopentyl-2-(4,6-dimethyl-3-oxo[1,2]thiazolo[5,4-b]pyridin-2(3H)-yl)acetamide
-
-
N-cyclopropyl-1-methyl-3-oxo-1,3-dihydro-2,1-benzothiazole-5-sulfonamide
-
-
N-ethylmaleimide
-
inhibits wild-type enzyme and deletion mutant H1[DELTA1-73]
N-hydroxyisoquinolinedione
-
N-hydroxypyridinedione
-
NEM
-
2 mM, 50% inhibition
nootkatin
-
i.e. 2-hydroxy-5-(3-methyl-2-butenyl)-4-(1-methylethyl)-2,4,6-cycloheptatrien-1-one
tropolone
-
i.e. 2-hydroxy-2,4,6-cycloheptatrien-1-one
Zn2+
-
in presence of Mg2+, inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
-
mitochondrial protein P32
slightly enhances the RNase H1 enzymatic activity
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000027 - 0.00003
D14R1D3:DNA18
-
0.000143 - 0.000158
RNA18:DNA18
-
0.0014
DNA/RNA hybrid
-
obtained by transcription of calf thymus DNA
-
0.0016
poly(rAdT)
-
-
-
0.000097 - 0.000106
RNA-DNA heteroduplex
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.158 - 0.4
D14R1D3:DNA18
-
0.983 - 3.48
RNA18:DNA18
-
0.025 - 3.5
RNA-DNA heteroduplex
-
additional information
additional information
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5 - 15
D14R1D3:DNA18
-
6.67 - 25
RNA18:DNA18
-
240 - 36000
RNA-DNA heteroduplex
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00019
2-(2,3-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00045
2-(2,5-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00833
2-(4,6-dimethyl-3-oxo-1,2-benzothiazol-2(3H)-yl)-N-propylacetamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.0005
2-(4-chlorophenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00005
2-(6-fluoro-3-oxo-1,2-benzothiazol-2(3H)-yl)-N-(4-methylphenyl)acetamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00002
2-phenyl-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.0019
2-[(cyclopentylcarbonyl)amino]-4-ethyl-5-methylthiophene-3-carboxamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.05
5-nitrofuran-2-carboxylic acid [[4-(4-bromophenyl)-thiazol-2-yl]-(tetrahydrofuran-2-ylmethyl)-carbamoyl] methyl ester
Homo sapiens
-
inhibitory activity against human RNase H1. IC50 value about 50 microM
0.0064
6-(naphthalen-2-yl)-3-(pyridin-3-yl)[1,2,4]triazolo[3,4-b][1,3,4]thiadiazole
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00001
6-fluoro-2-(2-methylphenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.0002
6-fluoro-2-(4-methylphenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.0054
N-(5-benzyl-1,3,4-thiadiazol-2-yl)-2-ethylhexanamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00048
N-benzyl-2-(6-fluoro-3-oxo-1,2-benzothiazol-2(3H)-yl)acetamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.00332
N-cyclopentyl-2-(4,6-dimethyl-3-oxo[1,2]thiazolo[5,4-b]pyridin-2(3H)-yl)acetamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
0.0004
N-cyclopropyl-1-methyl-3-oxo-1,3-dihydro-2,1-benzothiazole-5-sulfonamide
Homo sapiens
-
60 mM KCl, 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 0.01% (w/v) bovine serum albumin, 0.01% (v/v) Triton X-100, at 20°C
additional information
additional information
Homo sapiens
-
no inhibitory activity against human RNase H1 for 5-nitrofuran-2-carboxylic acid adamantan-1-carbamoyl methyl ester
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
-
assay at
7.9
-
assay at
8 - 8.5
-
in presence of 10 mM Mg2+
8.5 - 9
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 11
-
more than 60% activity at pH 6.5, about 70% activity at pH 7.0, maximum activity at pH 7.5, more than 80% activity between pH 8.0 and 11.0
7 - 10.5
-
pH 7.0: about 50% of maximal activity, pH 10.5: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21
-
assay at
24
-
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
rhabdomyosarcoma cell line, low level in enzyme type 1 and 2 activity
Manually annotated by BRENDA team
-
pancreatic carcinoma cell line, low level in enzyme type 1 activity, high level in enzyme type 2 activity
Manually annotated by BRENDA team
-
thymidine kinase 1-deficient osteosarcoma cell line
Manually annotated by BRENDA team
-
bladder carcinoma cell line, low level in enzyme type 1 activity, moderate level in enzyme type 2 activity
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
mutations in each of the three RNase H2 subunits are implicated in a human auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
physiological function
ribonuclease H2 is the major nuclear enzyme involved in the degradation of RNA/DNA hybrids and removal of ribonucleotides misincorporated in genomic DNA
evolution
-
reverse transcriptase (RT) and ribonuclease H are among the most ancient and abundant protein folds. RNases H may have evolved from ribozymes, related to viroids, early in the RNA world, forming ribosomes, RNA replicases and polymerases. Basic RNA-binding peptides enhance ribozyme catalysis. RT and ribozymes or RNases H are present today in bacterial group II introns, the precedents of transposable elements. Thousands of unique RTs and RNases H are present in eukaryotes, bacteria, and viruses
malfunction
metabolism
physiological function
additional information
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
RNH2A_HUMAN
299
0
33395
Swiss-Prot
other Location (Reliability: 1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33000
-
gel filtration
89000
-
sucrose density gradient centrifugation
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotrimer
arrangement of subunits to form an enzymatically active complex, structure of the heterotrimeric RNase H2 complex, overview
dimer
-
RNases H act as dimers, with two Mg2+ or other divalent cations being essential for correct protein structure, stability and enzyme activity
trimer
heterotrimer of subunits H2A, H2B, H2C, SDS-PAGE and gel filtration
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant subunts in a complex containing RNASEH2B residues 2-226, full-length RNASEH2C, and RNASEH2A with catalytic site mutations D34A and D169A, X-ray diffraction structure determination and analysis at 4.1-4.4 A resolution
analysis of the crystal structure of human RNase H1 in complex with a DNA/RNA duplex
-
crystal structure analysis, overview
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recombinant deletion mutant AB14-233C, sitting drop vapor diffusion method at 18°C, from 0.1 M MgCl2, 15% PEG 3350, 0.1 M Bis-Tris, pH 5.5, and 2 mM reduced glutathione, X-ray diffraction structure determination and analysis at 3.1 A resolution, molecular replacement
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the enzyme's hybrid binding domain complexed with a 12 bp RNA-DNA hybrid and an 6 bp RNA-DNA hybrid, molecular replacement, hanging drop vapour diffusion method, 21°C, 6 bp complex crystals are obtained with the well solution containing 10% PEG 3350, 0.2 M NaCl, 0.1 M Tris, pH 8.5, 12 bp complex crystals are obtained with 1.2 M NaCl and 0.1 M HEPES, pH 7.5, X-ray diffraction structure determination and anaylsis at 2.7-2.8 A and 2.1-2.2 A resolution, respectively
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A177T
naturally occuring mutation in subunit RNASEH2B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS. The mutation disrupts the interface between a RNASEH2B alpha-helix and the RNASEH2C kinked helix
D169A
active site residue mutation in subunit RNASEH2A
D34A
active site residue mutation in subunit RNASEH2A
K143I
naturally occuring mutation in subunit RNASEH2C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P138L
naturally occuring mutation in subunit RNASEH2C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P151S
naturally occuring mutation in subunit RNASEH2C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P76L
naturally occuring mutation in subunit RNASEH2C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R291H
naturally occuring mutation in subunit RNASEH2A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R69W
naturally occuring mutation in subunit RNASEH2C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
A177T
A185V
C147A
-
site-directed mutagenesis, mutant is active under reducing and oxidizing conditions
C147A/C148A
-
site-directed mutagenesis, mutant is active under reducing and oxidizing conditions
C148A
-
site-directed mutagenesis, mutant is active under reducing and oxidizing conditions
C191A
-
site-directed mutagenesis, mutant is active under reducing conditions, but shows poor activity under oxidizing conditions
D2Y/L3P
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
D39Y
-
naturally occuring mutation in subunit C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
F230L
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
G185V
mutation in subunit H2B associated with Aicardi-Goutieres' syndrome, near-normal activity
G83S
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
H264A
-
the mutation causes an about 100fold decrease in kcat under multiple-turnover conditions, but does not alter the Km value. The H264A mutant is not rescued by increasing the Mg2+ concentration to 80 mM
H86R
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
K143I
K162T
K59A/K60A
-
site-directed mutagenesis in the hybrid binding domain, the mutation abolishes dsRNA binding
L138F
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
L60R
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P138L
-
naturally occuring mutation in subunit C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P151S
-
naturally occuring mutation in subunit C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P43H
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
P76L
-
naturally occuring mutation in subunit C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R108W
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R13H
-
naturally occuring mutation in subunit C, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R186W
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R235Q
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R291H
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
R32A/R33A
-
site-directed mutagenesis in the hybrid binding domain outside of the interface, the mutation abolishes dsRNA binding
R35A
-
site-directed mutagenesis, the mutant shows a much lower specific activity than the wild-type enzyme
R57A
-
site-directed mutagenesis in the hybrid binding domain on the observed nucleic-acid interface, the mutation abolishes dsRNA binding
R72A/K73A
-
site-directed mutagenesis in the hybrid binding domain outside of the interface, the mutation abolishes dsRNA binding
S159I
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
S229P
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
T163I
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
T240M
-
naturally occuring mutation in subunit A, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
V142I
V183M
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
V185G
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
W43A
-
site-directed mutagenesis in the hybrid binding domain on the observed nucleic-acid interface, the mutation abolishes dsRNA binding, the mutant shows a much lower specific activity than the wild-type enzyme
W73L
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
Y219H
-
naturally occuring mutation in subunit B, the mutation is involved in auto-inflammatory disorder, Aicardi-Goutieres syndrome, AGS
Y29F
-
site-directed mutagenesis in the hybrid binding domain, the mutation abolishes dsRNA binding
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
partially purified enzyme extract is labile against freezing and dilution, addition of 45% glycerol
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
His-Trap Ni-NTA column chromatography
-
partially
-
recombinant C-terminally His6-tagged wild-type and mutant enzyme, without the predicted mitochondrial targeting sequence (residues 1-26), from insect Sf9 insect cells by nickel affinity and heparin affinity chromatography
recombinant enzyme
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged human ribonuclease H1 from Escherichia coli by nickel affinity chromatography
recombinant His-tagged RNase H1 from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography followed by reverse phase chromatography
-
recombinant RNase H1 from Escherichia coli strain BL21(DE3) using affinity and ion exchange chromatography
-
recombinant wild-type and mutant enzymes from Escherichia coli BL21(DE3)
-
RNase H1
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
simultaneous expression of GST-tagged RNASEH2B, and untagged RNASEH2A and RNASEH2C subunits from vector pGEX6P1 as polycistronic construct, in Escherichia coli strain Rosetta-2
expressed in Escherichia coli BL21(DE3)[pLysS] cells
-
expression in Escherichia coli and in HeLa cell
expression of enzyme type 1 and 2 as GFP-fusion proteins in all 6 cell lines, transfection of cell lines with antisense oligodeoxynucleotides directed against RNA polymerase II, replication protein A, and Ha-ras, determination of response in expression levels of the enzyme type 1 and 2, overview
-
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
expression of His-tagged RNase H1 in Escherichia coli strain BL21(DE3)
-
expression of human RNase H1 in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli BL21(DE3)
-
recombinant expression of C-terminally His6-tagged wild-type and mutant enzyme, without the predicted mitochondrial targeting sequence (residues 1-26), in Spodoptera frugiperda Sf9 insect cells using the baculovirus expression system
recombinant expression of His-tagged human ribonuclease H1 in Escherichia coli
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
molecular biology
-
the enzyme can be useful in regulation of RNA degradation in applications of technologies involving mRNA, method development, overview
additional information
RNA-directed off/on switch of RNase H activity using boronic ester formation
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Crouch, R.J.; Dirksen, M.L.
Ribonuclease H
Cold Spring Harbor Monogr. Ser.
14
211-254
1982
Bos taurus, Saccharomyces cerevisiae, Escherichia coli, Homo sapiens, Mus musculus, Rattus norvegicus, Xenopus laevis
-
Manually annotated by BRENDA team
Frank, P.; Albert, S.; Cazenave, C.; Toulme, J.J.
Purification and characterization of human ribonuclease HI
Nucleic Acids Res.
22
5247-5254
1994
Homo sapiens
Manually annotated by BRENDA team
Eder, P.S.; Walder, J.A.
Ribonuclease H from K562 human erythroleukemia cells. Purification, characterization, and substrate specificity
J. Biol. Chem.
266
6472-6479
1991
Homo sapiens
Manually annotated by BRENDA team
ten Asbroek, A.L.M.A.; Van Groenigen, M.; Nooij, M.; Baas, F.
The involvement of human ribonucleases H1 and H2 in the variation of response of cells to antisense phosphorothioate oligonucleotides
Eur. J. Biochem.
269
583-592
2002
Homo sapiens
Manually annotated by BRENDA team
Mangos, M.M.; Min, K.L.; Viazovkina, E.; Galarneau, A.; Elzagheid, M.I.; Parniak, M.A.; Damha, M.J.
Efficient RNase H-directed cleavage of RNA promoted by antisense DNA or 2'F-ANA constructs containing acyclic nucleotide inserts
J. Am. Chem. Soc.
125
654-661
2003
Escherichia coli, Homo sapiens
Manually annotated by BRENDA team
Lima, W.F.; Wu, H.; Nichols, J.G.; Manalili, S.M.; Drader, J.J.; Hofstadler, S.A.; Crooke, S.T.
Human RNase H1 activity is regulated by a unique redox switch formed between adjacent cysteines
J. Biol. Chem.
278
14906-14912
2003
Homo sapiens
Manually annotated by BRENDA team
Rydberg, B.; Game, J.
Excision of misincorporated ribonucleotides in DNA by RNase H (type 2) and FEN-1 in cell-free extracts
Proc. Natl. Acad. Sci. USA
99
16654-16659
2002
Saccharomyces cerevisiae, Escherichia coli, Homo sapiens, Pyrococcus furiosus
Manually annotated by BRENDA team
Nowotny, M.; Yang, W.
Stepwise analyses of metal ions in RNase H catalysis from substrate destabilization to product release
EMBO J.
25
1924-1933
2006
Homo sapiens, Halalkalibacterium halodurans (Q9KEI9)
Manually annotated by BRENDA team
Lima, W.F.; Nichols, J.G.; Wu, H.; Prakash, T.P.; Migawa, M.T.; Wyrzykiewicz, T.K.; Bhat, B.; Crooke, S.T.
Structural Requirements at the catalytic site of the heteroduplex substrate for human RNase H1 catalysis
J. Biol. Chem.
279
36317-36326
2004
Homo sapiens
Manually annotated by BRENDA team
Budihas, S.R.; Gorshkova, I.; Gaidamakov, S.; Wamiru, A.; Bona, M.K.; Parniak, M.A.; Crouch, R.J.; McMahon, J.B.; Beutler, J.A.; Le Grice, S.F.J.
Selective inhibition of HIV-1 reverse transcriptase-associated ribonuclease H activity by hydroxylated tropolones
Nucleic Acids Res.
33
1249-1256
2005
Escherichia coli, Homo sapiens
Manually annotated by BRENDA team
Tang, X.; Dmochowski, I.J.
Controlling RNA digestion by RNase H with a light-activated DNA hairpin
Angew. Chem.
45
3523-3526
2006
Homo sapiens
Manually annotated by BRENDA team
Patureau, B.M.; Hudson, R.H.; Damha, M.J.
Induction of RNase H activity by arabinose-peptide nucleic acid chimeras
Bioconjug. Chem.
18
421-430
2007
Homo sapiens
Manually annotated by BRENDA team
Lima, W.F.; Rose, J.B.; Nichols, J.G.; Wu, H.; Migawa, M.T.; Wyrzykiewicz, T.K.; Vasquez, G.; Swayze, E.E.; Crooke, S.T.
The positional influence of the helical geometry of the heteroduplex substrate on human RNase H1 catalysis
Mol. Pharmacol.
71
73-82
2007
Homo sapiens
Manually annotated by BRENDA team
Lima, W.F.; Rose, J.B.; Nichols, J.G.; Wu, H.; Migawa, M.T.; Wyrzykiewicz, T.K.; Siwkowski, A.M.; Crooke, S.T.
Human RNase H1 discriminates between subtle variations in the structure of the heteroduplex substrate
Mol. Pharmacol.
71
83-91
2007
Homo sapiens
Manually annotated by BRENDA team
Fuji, H.; Urano, E.; Futahashi, Y.; Hamatake, M.; Tatsumi, J.; Hoshino, T.; Morikawa, Y.; Yamamoto, N.; Komano, J.
Derivatives of 5-nitrofuran-2-carboxylic acid carbamoyl methyl ester inhibit RNase H activity associated with HIV-1 reverse transcriptase
J. Med. Chem.
52
1380-1387
2009
Homo sapiens
Manually annotated by BRENDA team
Fluiter, K.; Mook, O.R.; Vreijling, J.; Langkjaer, N.; Hojland, T.; Wengel, J.; Baas, F.
Filling the gap in LNA antisense oligo gapmers: the effects of unlocked nucleic acid (UNA) and 4-C-hydroxymethyl-DNA modifications on RNase H recruitment and efficacy of an LNA gapmer
Mol. Biosyst.
5
838-843
2009
Homo sapiens
Manually annotated by BRENDA team
Chon, H.; Vassilev, A.; DePamphilis, M.L.; Zhao, Y.; Zhang, J.; Burgers, P.M.; Crouch, R.J.; Cerritelli, S.M.
Contributions of the two accessory subunits, RNASEH2B and RNASEH2C, to the activity and properties of the human RNase H2 complex
Nucleic Acids Res.
37
96-110
2009
Homo sapiens, Homo sapiens (Q5TBB1), Homo sapiens (Q8TDP1)
Manually annotated by BRENDA team
Johnson, C.N.; Spring, A.M.; Sergueev, D.; Shaw, B.R.; Germann, M.W.
Structural basis of the RNase H1 activity on stereo regular borano phosphonate DNA/RNA hybrids
Biochemistry
50
3903-3912
2011
Homo sapiens
Manually annotated by BRENDA team
Ruhanen, H.; Ushakov, K.; Yasukawa, T.
Involvement of DNA ligase III and ribonuclease H1 in mitochondrial DNA replication in cultured human cells
Biochim. Biophys. Acta
1813
2000-2007
2011
Homo sapiens
Manually annotated by BRENDA team
Reijns, M.A.; Bubeck, D.; Gibson, L.C.; Graham, S.C.; Baillie, G.S.; Jones, E.Y.; Jackson, A.P.
The structure of the human RNase H2 complex defines key interaction interfaces relevant to enzyme function and human disease
J. Biol. Chem.
286
10530-10539
2011
Homo sapiens (O75792), Homo sapiens
Manually annotated by BRENDA team
Figiel, M.; Chon, H.; Cerritelli, S.M.; Cybulska, M.; Crouch, R.J.; Nowotny, M.
The structural and biochemical characterization of human RNase H2 complex reveals the molecular basis for substrate recognition and Aicardi-Goutieres syndrome defects
J. Biol. Chem.
286
10540-10550
2011
Homo sapiens
Manually annotated by BRENDA team
Liu, Y.; Xu, J.; Karimiahmadabadi, M.; Zhou, C.; Chattopadhyaya, J.
Synthesis of 2,4-propylene-bridged (carba-ENA) thymidine and its analogues: the engineering of electrostatic and steric effects at the bottom of the minor groove for nuclease and thermodynamic stabilities and elicitation of RNase H
J. Org. Chem.
75
7112-7128
2010
Homo sapiens
Manually annotated by BRENDA team
Beilhartz, G.; Goette, M.
HIV-1 ribonuclease H: structure, catalytic mechanism and inhibitors
Viruses
2
900-926
2010
Escherichia coli, Homo sapiens
Manually annotated by BRENDA team
Alla, N.R.; Nicholson, A.W.
Evidence for a dual functional role of a conserved histidine in RNA-DNA heteroduplex cleavage by human RNase H1
FEBS J.
279
4492-4500
2012
Homo sapiens
Manually annotated by BRENDA team
White, R.; Saxty, B.; Large, J.; Kettleborough, C.A.; Jackson, A.P.
Identification of small-molecule inhibitors of the ribonuclease H2 enzyme
J. Biomol. Screen.
18
610-620
2013
Homo sapiens
Manually annotated by BRENDA team
Zhou, Y.; Zhang, J.; Jiang, Q.; Lu, J.
An allosteric switch-based hairpin for label-free chemiluminescence detection of ribonuclease H activity and inhibitors
Analyst
144
1420-1425
2019
Homo sapiens (O60930)
Manually annotated by BRENDA team
Edwards, T.C.; Mani, N.; Dorsey, B.; Kakarla, R.; Rijnbrand, R.; Sofia, M.J.; Tavis, J.E.
Inhibition of HBV replication by N-hydroxyisoquinolinedione and N-hydroxypyridinedione ribonuclease H inhibitors
Antiviral Res.
164
70-80
2019
Hepatitis B virus, Homo sapiens (O60930), Homo sapiens
Manually annotated by BRENDA team
Moelling, K.; Broecker, F.; Russo, G.; Sunagawa, S.
RNase H as gene modifier, driver of evolution and antiviral defense
Front. Microbiol.
8
1745
2017
Saccharomyces cerevisiae, Homo sapiens, Mus musculus, Escherichia coli (P0A7Y4)
Manually annotated by BRENDA team
Liang, X.H.; Sun, H.; Nichols, J.G.; Crooke, S.T.
RNase H1-dependent antisense oligonucleotides are robustly active in directing RNA cleavage in both the cytoplasm and the nucleus
Mol. Ther.
25
2075-2092
2017
Homo sapiens (O60930)
Manually annotated by BRENDA team
Liang, X.H.; Nichols, J.G.; Sun, H.; Crooke, S.T.
Translation can affect the antisense activity of RNase H1-dependent oligonucleotides targeting mRNAs
Nucleic Acids Res.
46
293-313
2018
Homo sapiens (O60930)
Manually annotated by BRENDA team
Al-Behadili, A.; Uhler, J.P.; Berglund, A.K.; Peter, B.; Doimo, M.; Reyes, A.; Wanrooij, S.; Zeviani, M.; Falkenberg, M.
A two-nuclease pathway involving RNase H1 is required for primer removal at human mitochondrial OriL
Nucleic Acids Res.
46
9471-9483
2018
Homo sapiens (O60930), Homo sapiens
Manually annotated by BRENDA team
Reverte, M.; Barvik, I.; Vasseur, J.J.; Smietana, M.
RNA-directed off/on switch of RNase H activity using boronic ester formation
Org. Biomol. Chem.
15
8204-8210
2017
Homo sapiens (O60930)
Manually annotated by BRENDA team
Posse, V.; Al-Behadili, A.; Uhler, J.P.; Clausen, A.R.; Reyes, A.; Zeviani, M.; Falkenberg, M.; Gustafsson, C.M.
RNase H1 directs origin-specific initiation of DNA replication in human mitochondria
PLoS Genet.
15
e1007781
2019
Homo sapiens (O60930)
Manually annotated by BRENDA team