Any feedback?
Please rate this page
(all_enzymes.php)
(0/150)

BRENDA support

2.7.7.49: RNA-directed DNA polymerase

This is an abbreviated version!
For detailed information about RNA-directed DNA polymerase, go to the full flat file.

Word Map on EC 2.7.7.49

Reaction

a 2'-deoxyribonucleoside 5'-triphosphate
+
DNAn
=
diphosphate
 +
DNAn+1

Synonyms

Bst DNA polymerase, CS5 pol, DNA nucleotidyltransferase (RNA-directed), DNA polymerase, DNA polymerase I, FeLV RT, FIV RT, FV Pol, FV reverse transcriptase, Gag-Pol, HBV-pol, HIV reverse transcriptase, HIV-1 M RT, HIV-1 O RT, HIV-1 R, HIV-1 reverse transcriptase, HIV-1 RT, HIV-reverse transcriptase, HIV-RT, human hepatitis B virus polymerase, human immunodeficiency virus type 1 reverse transcriptase, iScript enzyme, K4 polymerase, K4PolI, M-MuLV reverse transcriptase, MMLV RT, Moloney Murine leukemia virus reverse transcriptase, Moloney murine leukemia virus RT, MoMLV RT, More, MuLV RT, MX162-RT, MX65-RT, nucleoside reverse transcriptase, nucleotidyltransferase, deoxyribonucleate, RNA-dependent, p66 RT, P72, PFV RT, Pol, polymerase/reverse transcriptase, prototype foamy virus reverse transcriptase, R2-RT, reverse transcriptase, reverse transcriptase/RNA dependent DNA polymerase, reverse-transcriptase, revertase, RNA dependent DNA polymerase, RNA revertase, RNA-dependent DNA polymerase, RNA-instructed DNA polymerase, RT, SFV RT, simian foamy virus reverse transcriptase, SS RT, SuperScript I reverse transcriptase, Superscript II, SUPERSCRIPT II reverse transcriptase, T. Z05 pol, telomerase, telomerase catalytic subunit, telomerase reverse transcriptase, TERT, Tgo-Pol, xenotropic murine leukemia virus-related virus reverse transcriptase, XMRV RT, XNA reverse transcriptase

ECTree

     2 Transferases
         2.7 Transferring phosphorus-containing groups
             2.7.7 Nucleotidyltransferases
                2.7.7.49 RNA-directed DNA polymerase

Engineering

Engineering on EC 2.7.7.49 - RNA-directed DNA polymerase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L180M
-
lamivudine-resistant mutant
M204V
-
lamivudine-resistant mutant
P306A
-
replication competency is reduced to 12.75% of the wild-type value
P306D
-
replication competency is increased to 121.57% of the wild-type value
P306E
-
replication competency is increased to 940.59% of the wild-type value
P306F
-
replication competency is reduced to 41.18% of the wild-type value
P306G
-
replication competency is reduced to 1.96% of the wild-type value
P306L
-
replication competency is increased to 128.04% of the wild-type value
P306R
-
replication competency is reduced to 20.26% of the wild-type value
P306S
-
replication competency is reduced to 47.25% of the wild-type value
P306T
-
replication competency is reduced to 4.5% of the wild-type value
P306V
-
replication competency is increased to 152.75% of the wild-type value
P306Y
-
replication competency is reduced to 10.98% of the wild-type value
A114G
-
mutant enzyme retains significant DNA polymerase activity. 5-6fold increase in the ability of the enzyme to discriminate against ddNTPs
A114S
-
mutant enzyme retains significant DNA polymerase activity, enzyme shows wild-type ddNTP/dNTP discrimination efficiency
A114T
-
mutant enzyme shows very low catalytic efficiency in nucleotide incorporation assays, due to the high KM-values for dNTP. Decreased susceptibility to AZTTP when poly(rA)/(dT)16 is used as substrate
A114V
-
mutant enzyme shows very low catalytic efficiency in nucleotide incorporation assays, due to the high KM-values for dNTP. Decreased susceptibility to AZTTP when poly(rA)/(dT)16 is used as substrate, enzyme shows wild-type ddNTP/dNTP discrimination efficiency
D67N/K70R/T215Y/K219Q
-
mutation results in a 1.5fold decrease in the rate constant for polymerization and a 2.5fold decrease in the equilibrium dissociation constant for 3'-azido-3'-deoxythymidine 5'-triphosphate compared to wild-type enzyme. These values translate into a 4fold decrease in selectivity for 3'-azido-3'-deoxythymidine 5'-monophosphate incorporation ba the mutant enzyme as compared to wild-type enzyme for RNA dependent DNA replication. No such decrease in selectivity is detected for DNA dependent replication
E478Q
-
polymerization activity of mutant E478Q that inactivates the RNase H catalytic site is much more sensitive to efavirenz than the wild type
G190V
mutant with high level of resistance to efavirenz and nevirapine
I132A
mutation confers low-level resistance to nevirapine and delavirdine
I132M
mutation confers high-level resistance to nevirapine and delavirdine (more than 10fold) and low-level resistance (about 2fold) to efavirenz
I135A
mutation confers high level resistance to nevirapine and delavirdine, but not to efavirenz
I135M
mutation confers low-level resistance (about 2fold) to nevirapine, delavirdine and efavirenz
K101E/G190S/M41lL/T215Y
-
patient isolate. Efavirenz stimulates the RNase H of an reverse transcriptase from a patient-derived virus that is highly resistant and grows more rapidly in the presence of low concentrations of efavirenz
K101R
non-nucleoside reverse transcriptase inhibitor resistance mutation
K103N
K103N/Y181C
-
mutation shows only moderate effects on either polymerase or RNAse H inhibitory potencies of GW8248 and TMC-125. IC50 for nevirapine is strongly enhanced. 1.3fold increase in IC50-value for capravirine, 21.8fold increase in IC50-value for efavirenz
K103R
non-nucleoside reverse transcriptase inhibitor resistance mutation
K65R/L74V
-
84fold decrease of the catalytic rate constant. Virus replication capacity is severely impaired relative to wild-type enzyme. Poor ability of K65R/L74V RT to use natural nucleotides relative to wild-type enzyme: 15% that of wild-type enzyme for dATP, 36% for dGTP, 50% for dTTP, and 25% for dCTP
K65R/V75I
-
a HIV-1 O RT variant that shows increased fidelity and stability compared to the wild-type enzyme
L74V
-
site-directed mutagenesis
M184V
M230D
non-nucleoside reverse transcriptase inhibitor resistance mutation
M230N
non-nucleoside reverse transcriptase inhibitor resistance mutation
N136A
mutant enzyme with very low reverse transcriptase activity
N137A
mutation confers no resistance to nevirapine, delavirdine and efavirenz
P140A
mutant enzyme with very low reverse transcriptase activity
Q151A
severe reduction in the polymerase activity withoput any significant effect on the affinity for dNTP substrate. The mutant is nearly devoid of diphosphorolytic activity on a RNA/primer-binding-site template-primer
Q151M
-
site-directed mutagenesis
Q151N
Q258C/C280S
the engineered reverse transcriptase protein (Q258C and C280S mutations) cross-linked to the dsDNA primer/template (20-mer/27-mer) containing the N2-cystamine 2'-deoxyguanosine six bases upstream from the priming site (P site) was used for the crystallographic studies
R78A
-
a HIV-1 O RT variant that shows increased fidelity and stability compared to the wild-type enzyme
T139A
mutant enzyme with very low reverse transcriptase activity
T139V
mutation confers no resistance to nevirapine, delavirdine and efavirenz
V148I
-
reduced dNTP binding affinity, mutation V148I disrupts positioning of Q151 for interaction with the 3'-OH of the incoming dTTP, mutant enzyme binds to AZTTP 18 times more tightly than to dTTP
V197I
non-nucleoside reverse transcriptase inhibitor resistance mutation
Y1152-naphthyl-Tyr
-
mutant of the p66 subunit of reverse transcriptase reconstituted with wild-type p51 subunit. Mutant enzyme inefficiently incorporates dCTP at low concentrations and is kinetically slower with all dCTP analogues tested. 5fold less efficient for dCTP incorporation and 15fold less efficient for 2',3'-dideoxy-CTP incorporation compared to the wild-type enzyme
Y115aminomethyl-Phe
-
mutant of the p66 subunit of reverse transcriptase reconstituted with wild-type p51 subunit. Mutant enzyme incorporates dCTP more efficiently than the wild-type and is resistant to the chain terminator (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate when examined in a steady-state fidelity assay. Mutant enzyme incorporates very little 3TCMP, even when the concentration of the chain terminator was 100times that of the dCTP
Y181C
Y188L
-
mutation shows only moderate effects on either polymerase or RNAse H inhibitory potencies of GW8248 and TMC-125. IC50 for nevirapine is strongly enhanced. 47.8fold increase in IC50-value for capravirine, 58fold increase in IC50-value for efavirenz
M184V
Q294A
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (12% increase of activity compared to the wild type enzyme)
Q294C
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (34% increase of activity compared to the wild type enzyme)
Q294E
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (11% increase of activity compared to the wild type enzyme)
Q294H
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (16% increase of activity compared to the wild type enzyme)
Q294M
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (4% increase of activity compared to the wild type enzyme)
Q294N
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (36% increase of activity compared to the wild type enzyme)
Q294P
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (13% increase of activity compared to the wild type enzyme)
Q294R
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (35% increase of activity compared to the wild type enzyme)
Q294S
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (19% increase of activity compared to the wild type enzyme)
Q294W
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (13% increase of activity compared to the wild type enzyme)
Q294Y
-
the mutation has no significant effect on the noticeable level of the DNA polymerase activity of the enzyme (3% increase of activity compared to the wild type enzyme)
A502V
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
A644V
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
D200N
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
D200N/L603W/T330P/L139P/E607K
-
site-directed mutagenesis, highly processive and thermostable multiply-mutated M-MuLV RT variant with 65fold improvement in comparison to the wild-type enzyme, the maximum temperature of the full-length cDNA synthesis is raised to 62°C, compared to 45°C for the wild-type enzyme
D524A
-
mutant exhibits similar dissociation constants for heteropolymeric DNA/DNA (2.9-6.5 nM) and RNA/DNA complexes (1.2-2.9 nM) like wild-type. Unlike the WT, the mutant enzymes is devoid of RNase H activity, and not able to degrade RNA in RNA/DNA complexes
E286R
-
mutant exhibits similar dissociation constants for heteropolymeric DNA/DNA (2.9-6.5 nM) and RNA/DNA complexes (1.2-2.9 nM) like wild-type. Unlike the WT, the mutant enzymes is devoid of RNase H activity, and not able to degrade RNA in RNA/DNA complexes
E286R/E302K/L435R/D524A
-
mutant exhibits similar dissociation constants for heteropolymeric DNA/DNA (2.9-6.5 nM) and RNA/DNA complexes (1.2-2.9 nM) like wild-type. Unlike the WT, the mutant enzymes is devoid of RNase H activity, and not able to degrade RNA in RNA/DNA complexes
E302K
-
mutant exhibits similar dissociation constants for heteropolymeric DNA/DNA (2.9-6.5 nM) and RNA/DNA complexes (1.2-2.9 nM) like wild-type. Unlike the WT, the mutant enzymes is devoid of RNase H activity, and not able to degrade RNA in RNA/DNA complexes
E607K
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
F155V
the mutation increase the affinity of the enzyme for ribonucleotides, without affecting the Vmax for catalysis, and thereby conferrs to the enzyme significant RNA polymerase activity
F625S
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
H126R
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
K103R
-
site-directed mutagenesis, the mutant is resistant to inhibition by tenofovir and 3'-azido-3'-deoxythymidine
K658R
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
L139P
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
L333Q
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
L435R
-
mutant exhibits similar dissociation constants for heteropolymeric DNA/DNA (2.9-6.5 nM) and RNA/DNA complexes (1.2-2.9 nM) like wild-type. Unlike the WT, the mutant enzymes is devoid of RNase H activity, and not able to degrade RNA in RNA/DNA complexes
L603W
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
L671P
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
M428L
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
N649S
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
P130S
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
P65S
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
Q190M
-
site-directed mutagenesis, the mutant is resistant to inhibition by tenofovir and 3'-azido-3'-deoxythymidine
Q84A
mutant enzyme displays higher DNA polymerase activities than wild-type enzyme. Vmax of Q84AH using dTTP as a substrate is about three times that of wild-type value, whereas the Km values are very similar
Q84A/F155V
Q84A mutation further improves RNA polymerase and DNA polymerase activity of mutant F155V
Q84N
mutant enzyme displays higher DNA polymerase activities than wild-type enzyme
T287A
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
T330P
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
W388R
-
site-directed mutagenesis, the mutant shows increased thermostability and -performance compared to the wild-type enzyme
Y344H
-
site-directed mutagenesis, the mutant shows decreased thermostability and -performance compared to the wild-type enzyme
D211N
-
still capable of in vitro polymerization, although it is blocked for in vivo transposition. D211N mutation has minimal effect on nucleotide binding but reduces the kpol by about 230fold. The mutation reduces binding affinity for both Mn2+ and Mg2+
F292A
-
slight diminuation in DNA-dependent DNA synthesis (under conditions allowing multiple rounds of synthesis). Significant reduction in polymerase activity in presence of heparin
F292Y
-
severly impaired Rnase H activity
G294A
-
mutation results in enhanced pausing at multiple positions of the DNA template in DNA-dependent DNA synthesis (under conditions allowing multiple rounds of synthesis). Significant reduction in polymerase activity in presence of heparin
Y298A
-
slight diminuation in DNA-dependent DNA synthesis (under conditions allowing multiple rounds of synthesis). Significant reduction in polymerase activity in presence of heparin
Y298W
-
significant reduction in polymerase activity in presence of heparin
F227L
-
mutant shows 2 to 3fold increase in infectivity
G211S
-
mutant shows 2 to 3fold increase in infectivity
I148V
-
mutant shows 2 to 3fold increase in infectivity
I257M
-
mutant shows 2 to 3fold increase in infectivity
K412E
-
mutant demonstrates a 12fold increase in infectivity compared to the parental virus SIVmneCl8 indicating a role for mutations in the connection domain of reverse transcriptase in influencing viral infectivity and replication
R173K
-
mutant shows 2 to 3fold increase in infectivity
T288A
-
mutant shows 2 to 3fold increase in infectivity
V108I
-
mutant shows 2 to 3fold increase in infectivity
F227L
-
mutant shows 2 to 3fold increase in infectivity
-
I257M
-
mutant shows 2 to 3fold increase in infectivity
-
K412E
-
mutant demonstrates a 12fold increase in infectivity compared to the parental virus SIVmneCl8 indicating a role for mutations in the connection domain of reverse transcriptase in influencing viral infectivity and replication
-
R173K
-
mutant shows 2 to 3fold increase in infectivity
-
V108I
-
mutant shows 2 to 3fold increase in infectivity
-
C232A
mutation in CP2 motif, modest defect in RNA binding
F230A
mutation in CP2 motif, no effect on RNA binding
H234A
mutation in CP2 motif, modest defect in RNA binding
I229A
mutation in CP2 motif, modest defect in RNA binding
R226A
mutation in CP2 motif, strong defect in RNA binding
R237A
mutation in CP2 motif, strong defect in RNA binding
Y231A
mutation in CP2 motif, no effect on RNA binding
D215A
-
enzyme mutant devoid of 3'-5' exonuclease activity
F388A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
L329A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
L329A/Q384A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
L329A/Y438A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
M408A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
Q384A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
Q384A/Y438A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
T326A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
Y438A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
F388A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
-
L329A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
-
Q384A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
-
T326A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
-
Y438A
-
site-directed mutagenesis, the mutant exhibits DNA-dependent and RNA-dependent DNA polymerase activities, while the wild-type enzyme is a DNA-depedent DNA polymerase
-
A608T/E520G/W827R
-
substitution rates per base for RNA-dependent DNA is similar or lower than that of avian myeloblastosis virus reverse transcriptase. Rate constants kcat of about two orders of magnitude larger than those of the Stoffel fragment
M747K/E742K
-
substitution rates per base for RNA-dependent DNA is similar to that of avian myeloblastosis virus reverse transcriptase
M761T/D547G/I584V
-
fidelity which is about 10-times higher than that of avian myeloblastosis virus reverse transcriptase. Almost tenfold improved catalytic efficiency as measured by the kcat/Km ratio when compared with the Stoffel fragment
N483Q/S486Q/T539N/Y545Q/D547T/P548Q/A570Q/D578Q/A597T/W604R/S612N/V730L/R736Q/S739N/M747R
-
selection of a polymerase with 15 mutations, mostly located at the template binding interface, by directed evolution of Thermus aquaticus DNA polymerase I, the mutant enzyme is a single variant of the Stoffel fragment of Taq DNA polymerase I, the enzyme shows broad template specificity and is a thermostable DNA-dependent and RNA-dependent DNA-polymerase, see also EC 2.7.7.7
K103R
site-directed mutagenesis, the mutant is resistant to inhibition by tenofovir and 3'-azido-3'-deoxythymidine
Q190M
site-directed mutagenesis, the mutant is resistant to inhibition by tenofovir and 3'-azido-3'-deoxythymidine
additional information