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Synonyms
reverse transcriptase, ribonuclease h, rnase h2, rnase hii, rnaseh2a, rnaseh1, ribonuclease hi, ribonuclease h2, rnase hiii, ribonuclease h1,
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DNA-RNA hybrid + H2O
ssDNA + 5'-phosphomonoester oligonucleotides
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in the pause of minus strang synthesis, RNAse H degrades the RNA template, with the exception of the polypurine tract sequence, immediately upstream of U3, which serves as a primer for plus-strand synthesis
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dsDNA oligonucleotide with a stretch of ribonucleotides + H2O
dsDNA oligonucleotide with 1 nt gap + 5'-monophosphate ribonucleotide
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enzyme excises misincorporated ribonucleotides in DNA
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dsDNA oligonucleotides with a single ribose + H2O
dsDNA oligonucleotides with 1 nt gap + 5'-monophosphate ribonucleotide
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preferred substrate, enzyme excises misincorporated ribonucleotides in DNA, enzyme places the first 5' nick, while the second 3' cut is made by Rad27p
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester + ?
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additional information
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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a mixture of oligonucleotides with 5'-phosphate termini and only a minor proportion of 5'-mononucleotide
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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T7 DNA-RNA hybrids
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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poly(rCdG)
mixture of oligonucleotides, ranging in size from dinucleotides to larger than hexanucleotides. No mononucleotides can be detected
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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M13 DNA:RNA[P*]DNA
oligoribonucleotides with 3'-hydroxyl and 5'-phosphate termini
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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poly(rUdA)
5'-phosphorylated oligonucleotides
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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poly(rCdI)
5'-phosphorylated oligonucleotides
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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poly(rAdT)
mixture of oligonucleotides, ranging in size from dinucleotides to larger than hexanucleotides. No mononucleotides can be detected
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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poly(rAdT)
5'-phosphorylated oligonucleotides
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RNA-DNA hybrid + H2O
ribonucleotide 5'-phosphomonoester
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RNase H2 incises the DNA 5'-of the ribonucleotide, generating DNA containing 3'-hydroxyl and 5'-phosphoribonucleotide ends
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additional information
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ribonuclease H(70) possesses cryptic reverse transcriptase activity
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additional information
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ribonuclease H(70) possesses cryptic reverse transcriptase activity
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additional information
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substrate specificity, enzyme plays a role in the repair of misincorporated ribonucleotides rather than or in addition to processing RNA*DNA hybrid molecules
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evolution
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the 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
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strains deficient in RNase H2 display a weak mutator phenotype which is consistent with a defect in DNA repair. RNase H2 defects cause alterations in the timing of cell cycle transitions
malfunction
disrupting the activity of the two enzymes RNase H1 and H2 (rnh1DELTA rnh201DELTA in Saccharomyces cerevisiae) is a useful tool for increasing the persistence of DNA:RNA hybrids and studying the effects of hybrid-induced instability. In the absence of RNase H activity, the levels of hybrids formed at susceptible loci increase dramatically. This increase in hybrids is associated with increased rates of genome instability that include loss of heterozygosity (LOH) events, loss of entire chromosomes, and recombination at the ribosomal locus. rnh1DELTA rnh201DELTA mutants display an increase in Rad52-GFP foci. Cells lacking RNase H1 and H2 have a larger fraction of persistent R-loop induced damage than wild-type cells or cells lacking only one of the RNases H, failure to observe accumulating foci early in the cell cycle, phenotype, overview
physiological function
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ribonucleotide excision repair is most efficient when the ribonucleotide is incised by RNase H2. RNase H1 fails to substitute for RNase H2 in the incision step of ribonucleotide excision repair
physiological function
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RNase H2 is implicated in the processing of the 5' ends of Okazaki fragments. RNase H2 also links DNA replication and DNA repair through ribonucleotide excision repair. The RNase H2 interaction network also functions to suppress genome instability
physiological function
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DNA replication requires RNA primers to initiate lagging strand DNA synthesis and their subsequent removal by the RNase. RNase H enzymes mediate viral and cellular replication and antiviral defense in eukaryotes and prokaryotes, splicing, R-loop resolvation, DNA repair. RNase H-like activities are also required for the activity of small regulatory RNAs. Virtually all known immune defense mechanisms against viruses, phages, transposable elements, and extracellular pathogens require RNase H-like enzymes. RNase H-like activities of retroviruses, transposable elements, and phages, have built up innate and adaptive immune systems throughout all domains of life
physiological function
R-loops are structures that form when RNA invades double-stranded DNA and hybridizes to complementary genomic sequences. R-loops can form spontaneously across many genomic loci, but the activity of two endogenous RNases H prevents their accumulation and persistence. RNase H enables efficient repair of R-loop induced DNA damage. RNase H1 and H2 are highly conserved ribonucleases with the ability to degrade the RNA moiety of a DNA:RNA hybrid. The RNases H are important protectors of genome stability, mechanisms, overview. The presence of either RNase H1 or H2 prevents the accumulation of DNA damage in G2-M
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G42S
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the mutant shows strongly reduced activity compared to the wild type enzyme
I183M
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site-directed mutagenesis in the polymerase domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
M520I
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
M520V
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
N398D
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
P45D/Y219A
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the mutant shows strongly reduced activity compared to the wild type enzyme
S392C
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
S442P
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
S469G
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
T467A
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site-directed mutagenesis in the connection domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
Y299C
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site-directed mutagenesis in the RNase H domain, the mutation results in a Mn2+ suppressor mutant less sensitive to Mn2+ inhibition
additional information
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construction of several deletion mutants
additional information
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generation of Ty1 reverse transcriptase/RNase H Mn2+ suppressor mutants capable of increased Ty1 transposition in pmr1DELTA cells for Mn2+ inhibition analysis, PMR1 codes for a P-type ATPase that regulates intracellular calcium and manganese ion homeostasis, and pmr1 mutants accumulate elevated intracellular manganese levels and display 100-fold less transposition than PMR1+ cells, suppressor point mutations localize not to the reverse transcriptase itself but to the RNase H domain of the protein, overview
additional information
generation of the rnh1DELTA rnh201DELTA mutant with disrupted activity of the two enzymes RNase H1 and H, kinetics of foci persistence in rnh1D rnh201D cells, phenotype, overview. Cell cycle delay is observed in the rnh1DELTA rnh201DELTA strain and in rnh1DELTA rnh201DELTA TOP1-AID cells. The Rad52-GFP foci in the rnh1DELTA rnh201DELTA double mutant accumulated in a window that begins at the boundary between S and G2-M. Screen for suppressors of hybrid-induced lethality, identifying Pif1-E467G, this allele suppresses auxin sensitivity when introduced into rnh1DELTA rnh201DELTA TOP1-AID cells, indicating its responsibility for the suppression of lethality in the mutant cells. Pif1 mutants inhibit break-induced replication. RPA190 mutants allow for repair of R-loop induced damage. Rpa190-K1482T and -V1486F suppress auxin sensitivity of rnh1DELTA rnh201DELTA TOP1-AID cells
additional information
generation of the rnh1DELTA rnh201DELTA mutant with disrupted activity of the two enzymes RNase H1 and H, kinetics of foci persistence in rnh1D rnh201D cells, phenotype, overview. Cell cycle delay is observed in the rnh1DELTA rnh201DELTA strain and in rnh1DELTA rnh201DELTA TOP1-AID cells. The Rad52-GFP foci in the rnh1DELTA rnh201DELTA double mutant accumulated in a window that begins at the boundary between S and G2-M. Screen for suppressors of hybrid-induced lethality, identifying Pif1-E467G, this allele suppresses auxin sensitivity when introduced into rnh1DELTA rnh201DELTA TOP1-AID cells, indicating its responsibility for the suppression of lethality in the mutant cells. Pif1 mutants inhibit break-induced replication. RPA190 mutants allow for repair of R-loop induced damage. Rpa190-K1482T and -V1486F suppress auxin sensitivity of rnh1DELTA rnh201DELTA TOP1-AID cells
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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
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brenda
Wyers, F.; Sentenac, A.; Fromageot, P.
Role of DNA-RNA hybrids in eukaryotes
Eur. J. Biochem.
69
377-383
1976
Saccharomyces cerevisiae
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brenda
Wyers, F.; Huet, J.; Sentenac, A.; Fromageot, P.
Role of DNA-RNA hybrids in eukaryotes. Characterization of yeast ribonuclease H1 and H2
Eur. J. Biochem.
69
385-395
1976
Saccharomyces cerevisiae
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brenda
Wintersberger, U.; Kuhne, C.; Karwan, R.
Three ribonucleases H and a reverse transcriptase from the yeast, Saccharomyces cerevisiae
Biochim. Biophys. Acta
951
322-329
1988
Saccharomyces cerevisiae
brenda
Cheriathundam, E.; Chang, L.M.S.
Purification and properties of two ribonuclease H enzymes from yeast
Arch. Biochem. Biophys.
219
110-120
1982
Saccharomyces cerevisiae
brenda
Karwan, R.; Kuehne, C.; Winterberger, U.
Ribonuclease H(70) from Saccharomyces cerevisiae possesses cryptic reverse transcriptase activity
Proc. Natl. Acad. Sci. USA
83
5919-5923
1986
Saccharomyces cerevisiae
brenda
Karwan, R.; Wintersberger, U.
Yeast ribonuclease H(70) cleaves RNA-DNA junctions
FEBS Lett.
206
189-192
1986
Saccharomyces cerevisiae
brenda
Karwan, R.; Wintersberger, U.
In addition to RNase H(70) two other proteins of Saccharomyces cerevisiae exhibit ribonuclease H activity
J. Biol. Chem.
263
14970-14977
1988
Saccharomyces cerevisiae
brenda
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
brenda
Yarrington, R.M.; Chen, J.; Bolton, E.C.; Boeke, J.D.
Mn2+ suppressor mutations and biochemical communication between Ty1 reverse transcriptase and RNase H domains
J. Virol.
81
9004-9012
2007
Saccharomyces cerevisiae
brenda
Allen-Soltero, S.; Martinez, S.L.; Putnam, C.D.; Kolodner, R.D.
A Saccharomyces cerevisiae RNase H2 interaction network functions to suppress genome instability
Mol. Cell. Biol.
34
1521-1534
2014
Saccharomyces cerevisiae
brenda
Sparks, J.L.; Chon, H.; Cerritelli, S.M.; Kunkel, T.A.; Johansson, E.; Crouch, R.J.; Burgers, P.M.
RNase H2-initiated ribonucleotide excision repair
Mol. Cell
47
980-986
2012
Saccharomyces cerevisiae
brenda
Chon, H.; Sparks, J.L.; Rychlik, M.; Nowotny, M.; Burgers, P.M.; Crouch, R.J.; Cerritelli, S.M.
RNase H2 roles in genome integrity revealed by unlinking its activities
Nucleic Acids Res.
41
3130-3143
2013
Saccharomyces cerevisiae, Saccharomyces cerevisiae W-303
brenda
Amon, J.D.; Koshland, D.
RNase H enables efficient repair of R-loop induced DNA damage
eLife
5
e20533
2016
Saccharomyces cerevisiae (P53942), Saccharomyces cerevisiae (Q04740)
brenda
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)
brenda