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Literature summary for 2.7.7.48 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
Search for more literature for 2.7.7.48

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli rhinovirus A16

Protein Variants

Protein Variants Comment Organism
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

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

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
K+ modest stimulation below 10 mM rhinovirus A16
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
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
rhinovirus A16
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 rhinovirus A16

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?
CTP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?
GTP + poly(C)
-
 rhinovirus A16 diphosphate + poly(C)n+1
-
 ?
GTP + poly(C)n
-
 rhinovirus A16 diphosphate + poly(C)n+1
-
 ?
GTP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?
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
-
 ?
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
-
 ?
nucleoside triphosphate + RNAn required for replication of the HRV RNA genome rhinovirus A16 diphosphate + RNAn+1
-
 ?
UTP + poly(A)n
-
 rhinovirus A16 diphosphate + poly(A)n+1
-
 ?
UTP + sshRNAn
-
 rhinovirus A16 diphosphate + sshRNAn+1
-
 ?

Subunits

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

Temperature Optimum [°C]

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

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
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]

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

pH Optimum

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

pH Range

pH Minimum pH Maximum Comment Organism
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

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