Protein Variants | Comment | Organism |
---|---|---|
D101N | mutation results in more than 95% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
D129A | Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 2.2fold higher than the wild-type value. The turnover number is 30% of the wild-type value | Archaeoglobus fulgidus |
D129N | mutation results in about 75% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 70% of the wild-type value. The turnover number is 20% of the wild-type value | Archaeoglobus fulgidus |
D167A | no loss of activity | Archaeoglobus fulgidus |
D37A | mutation results in about 90% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 80% of the wild-type value. The turnover number is 5% of the wild-type value | Archaeoglobus fulgidus |
D37N | mutation results in about 10% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 1.5fold higher than the wild-type value. The turnover number is 40% of the wild-type value | Archaeoglobus fulgidus |
D6N | mutation results in more than 95% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
E7Q | mutation results in more than 95% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
K143A | mutation results in about 80% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 31.3fold higher than the wild-type value. The turnover number is 150% of the wild-type value | Archaeoglobus fulgidus |
K39A | mutation results in about 30% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
L41A | no loss of activity | Archaeoglobus fulgidus |
additional information | of the highly conserved residues, mutations at Ser38, Lys39, Leu41, Arg45, Ser139, and Asp167 do not significantly affect the enzyme activity. Conversion of Asp6 to Asn, Glu7 to Gln, or Asp101 to Asn almost completely inactivated the enzyme, suggesting that these carboxylic acid groups of D6, E7, and D101 are essential for catalysis | Archaeoglobus fulgidus |
R146A | mutation results in more than 95% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 26.7fold higher than the wild-type value. The turnover number is 140% of the wild-type value | Archaeoglobus fulgidus |
R188A | no loss of activity | Archaeoglobus fulgidus |
R45A | mutation results in about 5% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
R46A | mutation results in about 50% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 60fold higher than the wild-type value. The turnover number is 130% of the wild-type value | Archaeoglobus fulgidus |
S139A | no loss of activity | Archaeoglobus fulgidus |
S139A | mutation results in about 40% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
S38A | mutation results in about 35% decrease in activity compared to wild-type enzyme | Archaeoglobus fulgidus |
Y164A | mutation results in more than 95% decrease in activity compared to wild-type enzyme. Km-value for the 8-mer RNA-DNA/DNA hybrid substrate is 44.7fold higher than the wild-type value. The turnover number is 160% of the wild-type value | Archaeoglobus fulgidus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Co2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Optimum concentration of Co2+ or Ni2+ needed for aRNase HII activity is 1 mM. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Co3+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Cu2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Mg2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Optimal enzyme activities in the presence of Mg2+ or Mn2+ are 3fold to 7fold higher than that with the other two metals. Maximum aRNase HII activity is observed at concentrations of 6.4 mM Mg2+. The specific activity determined in the presence of 50 mM Mn2+ is 35% of that determined in the presence of 6.4 mM Mg2+. When Mn2+ is added in the presence of 1.6 mM Mg2+, the enzyme activity increases gradually as the Mn2+ concentration reaches 50 mM and decreases after that point. At equal concentrations of Mn2+ and Mg2+ (1.6 mM), the enzyme activity is reduced 10-fold compared to the activity in the presence of only Mg2+. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Mn2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Optimal enzyme activities in the presence of Mg2+ or Mn2+ are 3fold to 7fold higher than that with the other two metals. Maximum aRNase HII activity is observed at concentrations of 50 mM Mn2+. The specific activity determined in the presence of 50 mM Mn2+ is 35% of that determined in the presence of 6.4 mM Mg2+. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Ni2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Optimum concentration of Co2+ or Ni2+ needed for aRNase HII activity is 1 mM. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus | |
Zn2+ | strictly metal-dependent nuclease. It exhibits activity in the presence of Mg2+, Mn2+, Co2+ or Ni2+, whereas no activity is observed in the absence of these metal ions. Little activity is detected in the presence of other metals including Co3+, Cu2+, Zn2+, and Ca2+ | Archaeoglobus fulgidus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Archaeoglobus fulgidus | O29634 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
RNA/DNA hybrid + H2O | model Okazaki fragment 18-mer RNA-DNA/DNA substrate (Q18), RNase H is a structure-specific endonuclease, it cleaves the 25-bp RNA/DNA hybrid at multiple sites, indicating that the enzyme cleaves RNA/DNA in a sequence-independent manner. In the absence of complementary DNA, the chimeric RNA-DNA strand is not cleaved by the enzyme | Archaeoglobus fulgidus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
aRNase HII | - |
Archaeoglobus fulgidus |
RNase HII | - |
Archaeoglobus fulgidus |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme is involved in RNA primer removal during DNA replication | Archaeoglobus fulgidus |