Activating Compound | Comment | Organism | Structure |
---|---|---|---|
dithiothreitol | 10 mM, required to achieve maximal polymerase activity | Zika virus |
Cloned (Comment) | Organism |
---|---|
a codon-optimized cDNA sequence of ZIKV NS5 (amino acids 2522 to 3423; strain H/PF/2013) is synthesized de novo. The ZIKV NS5 polymerase portion is PCR amplified and cloned into a pMal-c5X vector under tac promoter control. Expression in Escherichia coli | Zika virus |
expression in Escherichia coli | Dengue virus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | active-site mutation, GDD -> GAA, in amino acid positions 663 to 665 of NS5 causes the complete loss of NS5 polymerase activity | Zika virus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3'-dATP | incorporation of the 3'-deoxynucleotide triphosphate fully prevents extension of a product in the presence of the next correct ribonucleotide | Zika virus | |
3'-dCTP | incorporation of the 3'-deoxynucleotide triphosphate fully prevents extension of a product in the presence of the next correct ribonucleotide | Zika virus | |
3'-dGTP | incorporation of the 3'-deoxynucleotide triphosphate fully prevents extension of a product in the presence of the next correct ribonucleotide | Zika virus | |
3'-dUTP | incorporation of the 3'-deoxynucleotide triphosphate fully preventes extension of a product in the presence of the next correct ribonucleotide | Dengue virus | |
3'-dUTP | incorporation of the 3'-deoxynucleotide triphosphate fully prevents extension of a product in the presence of the next correct ribonucleotide | Zika virus | |
4'-C-azidocytidine-5'-triphosphate | causes only partial chain termination | Zika virus | |
beta-D-2'-alpha-deoxy-2'-beta-fluoro-2'-C-methyluridine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Dengue virus | |
beta-D-2'-alpha-deoxy-2'-beta-fluoro-2'-C-methyluridine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-beta-C-ethynyl-7-deaza-adenosine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-beta-C-ethynyluridine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Dengue virus | |
beta-D-2'-beta-C-ethynyluridine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-beta-C-methyladenosine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-beta-C-methylcytidine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-beta-C-methylguanosine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-beta-C-methyluridine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Dengue virus | |
beta-D-2'-beta-C-methyluridine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-deoxy-2'-alpha-fluoro-2'-beta-C-methylcytidine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
beta-D-2'-deoxy-2'-alpha-fluoro-2'-beta-C-methylguanosine-5'-triphosphate | causes chain termination after incorporation into nascent RNA | Zika virus | |
imidazole | tolerable at low concentrations, but inhibits at concentrations higher than 25 mM | Zika virus | |
NaCl | tolerable at low concentrations, but inhibits at concentrations higher than 25 mM | Zika virus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | Mn2+x02is required for enzymatic activity, while Mg2+x02 is not | Zika virus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dengue virus | - |
- |
- |
Zika virus | A0A024B7W1 | genome polyprotein | - |
Purification (Comment) | Organism |
---|---|
- |
Dengue virus |
- |
Zika virus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
nucleoside triphosphate + RNAn | - |
Dengue virus | diphosphate + RNAn+1 | - |
? | |
nucleoside triphosphate + RNAn | DNA can serve as a template for ZIKV NS5 with an efficiency similar to that of an RNA template. The enzyme (NS5) can utilize single-stranded DNA but not double-stranded DNA as a template or a primer to synthesize RNA. Both full-length NS5 and a truncated NS5 containing the polymerase domain can carry out in vitro RNA-dependent RNA synthesis | Zika virus | diphosphate + RNAn+1 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NS5 | - |
Dengue virus |
NS5 | - |
Zika virus |
RNA-dependent RNA polymerase | - |
Dengue virus |
RNA-dependent RNA polymerase | - |
Zika virus |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.00033 | - |
pH 7.5, 35°C | Dengue virus | beta-D-2'-beta-C-ethynyluridine-5'-triphosphate | |
0.00046 | - |
pH 7.5, 35°C, measured in the presence of 0.001 mM competing UTP, RNA template | Zika virus | beta-D-2'-beta-C-ethynyluridine-5'-triphosphate | |
0.00067 | - |
pH 7.5, 35°C, measured in the presence of 0.001 mM competing UTP, RNA template | Zika virus | 3'-dUTP | |
0.00068 | - |
pH 7.5, 35°C | Dengue virus | 3'-dUTP | |
0.00494 | - |
pH 7.5, 35°C | Dengue virus | beta-D-2'-beta-C-methyluridine-5'-triphosphate | |
0.00578 | - |
pH 7.5, 35°C, measured in the presence of 0.001 mM competing UTP, RNA template | Zika virus | beta-D-2'-beta-C-methyluridine-5'-triphosphate | |
0.05513 | - |
pH 7.5, 35°C | Dengue virus | beta-D-2'-alpha-deoxy-2'-beta-fluoro-2'-C-methyluridine-5'-triphosphate | |
0.09076 | - |
pH 7.5, 35°C, measured in the presence of 0.001 mM competing UTP, RNA template | Zika virus | beta-D-2'-alpha-deoxy-2'-beta-fluoro-2'-C-methyluridine-5'-triphosphate |