Cloned (Comment) | Organism |
---|---|
gene rnc, expression of wild-type and mutant enzymes in strain BL21(DE3) as His-tagged enzymes | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a mutant enzyme RNase III[DELTAdsRBD] lacking the dsRNA binding domain, the mutant is still catalytically active at low salt concentrations in presence of either 25 mM Mg2+ or 5 mM Mn2+ with slightly reduced catalytic efficiency, but unaltered substrate specificity | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ethidium bromide | inhibition of wild-type and deletion mutant, intercalative mode of inhibition, competitive, 50% inhibition at 0.01 mM | Escherichia coli | |
Mn2+ | inhibition of wild-type and deletion mutant at high concentration due to metal ion occupancy at an inhibitory site of the enzyme, the deletion mutant is less sensitive than the wild-type enzyme | Escherichia coli | |
additional information | enzyme is inhibited by Watson-Crick bp antideterminants, overview | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00011 | - |
R1.1 RNA | recombinant wild-type enzyme, 37°C, in presence of 1 mM Mn2+ | Escherichia coli | |
0.00027 | - |
R1.1 RNA | recombinant enzyme mutant RNase III[DELTAdsRBD], 37°C, in presence of 5 mM Mn2+ | Escherichia coli | |
0.00034 | - |
R1.1 RNA | recombinant wild-type enzyme, 37°C, in presence of 5 mM Mg2+ | Escherichia coli | |
0.003 | - |
R1.1 RNA | recombinant enzyme mutant RNase III[DELTAdsRBD], 37°C, in presence of 25 mM Mg2+ | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | dependent on, optimal at 25 mM | Escherichia coli | |
Mn2+ | can substitute for Mg2+ in vitro, at 5 mM | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dsRNA + H2O | Escherichia coli | enzyme plays a key role in diverse maturation and degradation processes | mature RNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Endonucleolytic cleavage to a 5'-phosphomonoester | Glu117 is essential for phosphodiester hydrolysis but not for substrate binding, the dsRNA-binding domain is important for substrate binding but not for catalytic activity, while the catalytic domain is important for catalytic activity but not for substrate binding | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dsRNA + H2O | enzyme plays a key role in diverse maturation and degradation processes | Escherichia coli | mature RNA | - |
? | |
dsRNA + H2O | double-strand RNA-specific, the dsRNA-binding domain is important for substrate binding but not for catalytic activity, while the catalytic domain is important for catalytic activity but not for substrate binding | Escherichia coli | mature RNA | - |
? | |
additional information | no activity with (rA)25, (rU)25, (rC)25, dsDNA, ssDNA and an RNA-DNA hybrid | Escherichia coli | ? | - |
? | |
R1.1 RNA + H2O | substrate is enzymatically synthesized based on the R1.1 processing signal, which is encoded in the phage T7 genetic early region between genes 1.0 and 1.1, 1 cleavage site | Escherichia coli | 2 fragments of R1.1 RNA | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme contains a conserved dsRNA-binding and a conserved catalytic domain | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
More | enzyme belongs to the ribonuclease III superfamily | Escherichia coli |
RNase III | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.023 | - |
R1.1 RNA | recombinant wild-type enzyme, 37°C, in presence of 1 mM Mn2+ | Escherichia coli | |
0.048 | - |
R1.1 RNA | recombinant enzyme mutant RNase III[DELTAdsRBD], 37°C, in presence of 5 mM Mn2+ | Escherichia coli | |
0.052 | - |
R1.1 RNA | recombinant enzyme mutant RNase III[DELTAdsRBD], 37°C, in presence of 25 mM Mg2+ | Escherichia coli | |
0.063 | - |
R1.1 RNA | recombinant wild-type enzyme, 37°C, in presence of 5 mM Mg2+ | Escherichia coli |