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Literature summary for 3.1.13.2 extracted from

  • Lawler, J.L.; Mukherjee, P.; Coen, D.M.
    Herpes simplex virus 1 DNA polymerase RNase H activity acts in a 3'-to-5' direction and is dependent on the 3'-to-5' exonuclease active site (2018), J. Virol., 92, e01813-17 .
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
medicine HSV-1 DNA polymerase is a crucial target for antivirals against HSV-1 infection Human alphaherpesvirus 1

Protein Variants

Protein Variants Comment Organism
additional information generation of a 3'-to-5' exonuclease-deficient mutant, D368A Pol. Wild-type and D368A mutant DNA polymerase exhibit similar polymerase activities, but the mutant enzyme is drastically impaired for 3'-to-5' exonuclease activity, with no activity detected even at high enzyme-to-DNA substrate ratios. The mutant shows no detectable ability to excise RNA with either a 3' or 5' terminus in contrast to the wild-type enzyme. Wild-type HSV Pol exhibits readily detectable RNase H activity on 6-FAM-labeled hairpin RNA-DNA substrate with a 3' RNA terminus in the 3'-to-5' direction, while the mutant is inactive. Neither wild-type nor D368A Pol exhibits detectable RNase H activity on a substrate with a 5' RNA terminus Human alphaherpesvirus 1

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Human alphaherpesvirus 1

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Human alphaherpesvirus 1 Herpes simplex virus 1 DNA polymerase shows also RNase H activity and acts in a 3'-to-5' direction, the activity is dependent on the 3'-to-5' exonuclease active site. No RNase H activity of HSV-1 DNA polymerase on RNA-DNA hybrids with 5' RNA termini ?
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Organism

Organism UniProt Comment Textmining
Human alphaherpesvirus 1
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HSV-1
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Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
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reaction velocities of wild-type and mutant enzymes in polymerase (EC 2.7.7.49) and 3'-to-5' RNase H activity, overview. Comparison of rates of 5'-to-3' polymerase, 3'-to-5' exonuclease, and degradation of RNA-DNA with 3' termini over a range of enzyme concentrations. The initial rates of the wild-type 5'-to-3' polymerase, D368A 5'-to-3' polymerase, wild-type 3'-to-5' exonuclease, and wild-type degradation of RNA with 3' termini in RNA-DNA hybrids are determined Human alphaherpesvirus 1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information Herpes simplex virus 1 DNA polymerase shows also RNase H activity and acts in a 3'-to-5' direction, the activity is dependent on the 3'-to-5' exonuclease active site. No RNase H activity of HSV-1 DNA polymerase on RNA-DNA hybrids with 5' RNA termini Human alphaherpesvirus 1 ?
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additional information in vitro assays are performed utilizing purified wild-type Pol and the D368A exonuclease-deficient mutant, testing the ability of these enzymes to extend a fluorescently labeled DNA hairpin primer-template and degrade dsDNA and RNA-DNA hybrid hairpin substrates over time, assay of RNase H activity using differentially end-labeled templates with 5' or 3' RNA termini, e.g. 6-FAM-labeled hairpin RNA-DNA substrate with a 3' RNA terminus. Wild-type HSV Pol exhibits readily detectable RNase H activity on this substrate in the 3'-to-5' direction, while the mutant is inactive. Neither wild-type nor D368A Pol exhibits detectable RNase H activity on a substrate with a 5' RNA terminus Human alphaherpesvirus 1 ?
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Synonyms

Synonyms Comment Organism
3'-to-5' RNase H
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Human alphaherpesvirus 1
More cf. EC 2.7.7.49 Human alphaherpesvirus 1
ribonuclease H
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Human alphaherpesvirus 1
RNase H
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Human alphaherpesvirus 1

General Information

General Information Comment Organism
additional information HSV-1 Pol contains four domains: the pre-NH2-terminal domain, the NH2-terminal domain, the 3'-to-5' exonuclease (Exo) domain, and the polymerase domain, composed of the palm, finger, and thumb subdomains Human alphaherpesvirus 1
physiological function among the activities of herpes simplex virus 1 DNA polymerase (HSV-1 Pol) is an intrinsic RNase H activity that cleaves RNA from RNA-DNA hybrids. The wild-type HSV-1 Pol is able to cleave RNA from the annealed RNA-DNA hairpin template, but only detectably with a 3' RNA terminus in a 3'-to-5' direction and at a rate lower than that of the exonuclease activity. These results suggest that HSV-1 Pol does not have an RNase H separable from its 3'-to-5' exonuclease activity and that this activity prefers DNA degradation over degradation of RNA from RNA-DNA hybrids. No RNase H activity of HSV-1 DNA polymerase on RNA-DNA hybrids with 5' RNA termini Human alphaherpesvirus 1