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Literature summary extracted from

  • Hung, M.; Gibbs, C.S.; Tsiang, M.
    Biochemical characterization of Rhinovirus RNA-dependent RNA polymerase (2002), Antiviral Res., 56, 99-114.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.48 expression in Escherichia coli rhinovirus A16

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.48 additional information the activity of the 6His-tagged enzyme HRV-16 3D polymerase is identical to HRV-16 3D polymerase without the 6His-tag rhinovirus A16

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.48 ATP IC50: 0.0053 mM rhinovirus A16
2.7.7.48 CTP IC50: 0.0015 mM rhinovirus A16
2.7.7.48 dATP IC50 above 0.5 mM rhinovirus A16
2.7.7.48 dCTP IC50: 0.04 mM rhinovirus A16
2.7.7.48 ddATP IC50 above 1 mM rhinovirus A16
2.7.7.48 ddCTP IC50 above 1 mM rhinovirus A16
2.7.7.48 ddGTP IC50: above 0.2 mM rhinovirus A16
2.7.7.48 ddTTP IC50 above 0.5 mM rhinovirus A16
2.7.7.48 dGTP IC50: 0.1 mM rhinovirus A16
2.7.7.48 dTTP IC50 above 0.5 mM rhinovirus A16
2.7.7.48 gliotoxin IC50: 0.15 mM rhinovirus A16
2.7.7.48 GTP IC50: 0.0002 mM rhinovirus A16
2.7.7.48 K+ above 10 mM rhinovirus A16
2.7.7.48 Mn2+ polymerase activity drops sharply at 3 mM rhinovirus A16
2.7.7.48 additional information the enzyme is resistant to 0.02 mg/ml rifampicin rhinovirus A16
2.7.7.48 UTP IC50: 0.0023 mM rhinovirus A16
2.7.7.48 Zn2+ IC50: 0.0006 mM, suppression of inhibition by dithiothreitol rhinovirus A16

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.48 0.00026
-
 CTP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.001
-
 GTP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.0021
-
 UTP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.007
-
 poly(C)n pH 7.0, 30°C, poly(C) as template rhinovirus A16
2.7.7.48 0.008
-
 ATP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.012
-
 poly(A)n pH 7.0, 30°C, poly(A) as template rhinovirus A16

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.48 K+ modest stimulation below 10 mM rhinovirus A16
2.7.7.48 Mg2+ in presence of Mg2+ significant activity is observed when poly(A) or poly(C) is used as template and the activity is template and primer-dependent. Poly(G) and poly(U) templates are not efficient substrates. Biotinylated oligoDNA primers appear to work slightly more efficiently than oligoRNA primers. Divalent cation required, optimal concentration for Mg2+ is 1 mM rhinovirus A16
2.7.7.48 Mn2+ in presence of Mn2+ activity is stimulates by 2.5-5.6fold. RNA synthesis using poly(C) as template becomes primer-independent, about 2.5fold stimulation at 1 mM rhinovirus A16

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.48 54000
-
 x * 54000, SDS-PAGE rhinovirus A16

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.48 nucleoside triphosphate + RNAn rhinovirus A16 required for replication of the HRV RNA genome diphosphate + RNAn+1
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.48 rhinovirus A16
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.48
-
 rhinovirus A16

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.48 ATP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?
2.7.7.48 CTP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?
2.7.7.48 GTP + poly(C)
-
 rhinovirus A16 diphosphate + poly(C)n+1
-
 ?
2.7.7.48 GTP + poly(C)n
-
 rhinovirus A16 diphosphate + poly(C)n+1
-
 ?
2.7.7.48 GTP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?
2.7.7.48 nucleoside triphosphate + RNAn in presence of Mg2+ significant activity is observed when poly(A) or poly(C) is used as template and the activity is template and primer-dependent. Poly(G) and poly(U) templates are not efficient substrates. Biotinylated oligoDNA primers appear to work slightly more efficiently than oligoRNA primers. In presence of Mn2+ activity is stimulated 2.5-5.6fold. RNA synthesis using poly(C) as template becomes primer-independent rhinovirus A16 diphosphate + RNAn+1
-
 ?
2.7.7.48 nucleoside triphosphate + RNAn RNA polymerase activity on homopolymeric templates poly(A) and poly(C) and heteropolymeric RNA templates primed with either RNA or DNA oligonucleotide primers or self-primed by a copy-back mechanism rhinovirus A16 diphosphate + RNAn+1
-
 ?
2.7.7.48 nucleoside triphosphate + RNAn required for replication of the HRV RNA genome rhinovirus A16 diphosphate + RNAn+1
-
 ?
2.7.7.48 UTP + poly(A)n
-
 rhinovirus A16 diphosphate + poly(A)n+1
-
 ?
2.7.7.48 UTP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.7.48 ? x * 54000, SDS-PAGE rhinovirus A16

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.48 30
-
 poly(A) as template and biotinylated oligo(dU)15 as the primer rhinovirus A16

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.48 17 37 17°C: about 20% of maximal activity, dramatic drecrease in activity between 32°C and 37°C, 37°C: no activity, poly(A) as template and biotinylated oligo(dU)15 as the primer rhinovirus A16

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.48 0.000505
-
 CTP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.000617
-
 ATP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.00375
-
 GTP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.00478
-
 UTP pH 7.0, 30°C, sshRNA as template rhinovirus A16
2.7.7.48 0.0211
-
 GTP pH 7.0, 30°C, poly(C) as template rhinovirus A16
2.7.7.48 0.23
-
 UTP pH 7.0, 30°C, poly(A) as template rhinovirus A16

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.48 7.3
-
 poly(A) as template and biotinylated oligo(dU)15 as the primer rhinovirus A16

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.48 6.5 8 pH 6.5: about 50% of maximal activity, pH 9.0: about 55% of maximal activity, poly(A) as template and biotinylated oligo(dU)15 as the primer rhinovirus A16

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.7.7.48 0.0002
-
 IC50: 0.0002 mM rhinovirus A16 GTP
2.7.7.48 0.0006
-
 IC50: 0.0006 mM, suppression of inhibition by dithiothreitol rhinovirus A16 Zn2+
2.7.7.48 0.0015
-
 IC50: 0.0015 mM rhinovirus A16 CTP
2.7.7.48 0.0023
-
 IC50: 0.0023 mM rhinovirus A16 UTP
2.7.7.48 0.0053
-
 IC50: 0.0053 mM rhinovirus A16 ATP
2.7.7.48 0.04
-
 IC50: 0.04 mM rhinovirus A16 dCTP
2.7.7.48 0.1
-
 IC50: 0.1 mM rhinovirus A16 dGTP
2.7.7.48 0.15
-
 IC50: 0.15 mM rhinovirus A16 gliotoxin
2.7.7.48 0.2
-
 IC50: above 0.2 mM rhinovirus A16 ddGTP
2.7.7.48 0.5
-
 IC50 above 0.5 mM rhinovirus A16 dATP
2.7.7.48 0.5
-
 IC50 above 0.5 mM rhinovirus A16 dTTP
2.7.7.48 0.5
-
 IC50 above 0.5 mM rhinovirus A16 ddTTP
2.7.7.48 1
-
 IC50 above 1 mM rhinovirus A16 ddCTP
2.7.7.48 1
-
 IC50 above 1 mM rhinovirus A16 ddATP