Application | Comment | Organism |
---|---|---|
medicine | HSV-1 DNA polymerase is a crucial target for antivirals against HSV-1 infection | Human alphaherpesvirus 1 |
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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Human alphaherpesvirus 1 |
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 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human alphaherpesvirus 1 | - |
HSV-1 | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
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 | 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 | ? | - |
? | |
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 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3'-to-5' RNase H | - |
Human alphaherpesvirus 1 |
More | cf. EC 2.7.7.49 | Human alphaherpesvirus 1 |
ribonuclease H | - |
Human alphaherpesvirus 1 |
RNase H | - |
Human alphaherpesvirus 1 |
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 |