2.7.7.7: DNA-directed DNA polymerase

mutation in the spacer region of the alpha-subunit. Obtained in small amounts due to low solubility, mutant has barely detectable DNA polymerase activity

DELTA483533

expressed efficiently and purified as soluble holoenzyme complex associated with wild-type beta-subunit. The DNA polymerase activity of the mutant holoenzyme is reduced greatly as compared to wild type

DELTA536581

expressed efficiently and purified as soluble holoenzyme complex associated with wild-type beta-subunit. The DNA polymerase activity of the mutant enzyme is 80% of wild-type holoenzyme

DELTA666742

mutation in the spacer region of the alpha-subunit. Obtained in small amounts due to low solubility, mutant has barely detectable DNA polymerase activity

F576A

mutation in alpha-subunit, mutant enzyme retains about 50% of wild-type activity

G575A

mutation in alpha-subunit, mutant enzyme has nearly normal activity

G575A/W576A/F578A

mutation in alpha-subunit completely reduces DNA polymerase activity

K557A

mutation in alpha-subunit, mutant enzyme has nearly normal activity

K687A/D688A/F689A

mutation in alpha-subunit resulted in DNA polymerase activities that is 6080% of wild-type enzyme

L558A

mutation in alpha-subunit, mutant enzyme retains about 50% of wild-type activity

P556A

mutation in alpha-subunit, mutant enzyme has nearly normal activity

P556A/K557A/L558A

mutation in alpha-subunit completely reduces DNA polymerase activity

S719A/Y720A/W721

mutation in alpha-subunit completely reduces DNA polymerase activity

W576A

mutation in alpha-subunit, mutant enzyme is nearly inactive

Y419A/E420A/D421A

mutation in alpha-subunit resulted in DNA polymerase activities that is 6080% of wild-type enzyme

F12A

mutant enzyme shows disproportionately reduced activity on the damaged template

676145

F13V

mutant enzyme is almost unable to carry out translesion synthesis over N2-furfuryl-dG, although its activity on undamaged DNA is unaffected. The F13V mutation has a modest effect on the ability of DinB to discriminate against ribonucleotides, increasing the frequency of their misincorporation from less than 0.00001 to 0.001

676145

F771A

the mutation shows 30% decreased polymerase activity compared to the wild type enzyme

L823A

the mutation shows 3% decreased polymerase activity compared to the wild type enzyme

N678A

no change in polymerase activity, increased mismatch-directed exonuclease activity

P680G

reduced kcat, no change in relative DNA binding affinity or Km, nearly complete loss in the processive mode of DNA synthesis

P680Q

reduced kcat, no change in relative DNA binding affinity or Km, nearly complete loss in the processive mode of DNA synthesis

Q667A

polymerase defective, no change in exonuclease activity

R821A

the mutation shows 19% increased polymerase activity compared to the wild type enzyme

R822A

the mutation shows 5% increased polymerase activity compared to the wild type enzyme

R822A/Y824A

the mutation shows 36% increased polymerase activity compared to the wild type enzyme

Y824A

the mutation shows 25% increased polymerase activity compared to the wild type enzyme

L561A

the mutant has pre-steadystate Kd,app and kpol values for the incorporation of correct dNMPs that are comparable to or exhibits kpol values that are somewhat greater than wild type DNA polymerase

L561A/Y567A

the mutant has pre-steadystate Kd,app and kpol values for the incorporation of correct dNMPs that are comparable to or exhibits kpol values that are somewhat greater than wild type DNA polymerase

L561A/Y567A/S565G

the mutant has pre-steadystate Kd,app and kpol values for the incorporation of correct dNMPs that are comparable to or exhibits kpol values that are somewhat greater than wild type RB69 DNA polymerase

Y567A

the mutant has pre-steadystate Kd,app and kpol values for the incorporation of correct dNMPs that are comparable to or exhibits kpol values that are somewhat greater than wild type DNA polymerase

D424A

Geobacillus anatolicus

a proofreading deficient mutant of the Klenow fragment

721761

D378A

site-directed mutagenesis, the mutant is inactive in presence of Mg2+

D378E

site-directed mutagenesis, the mutant is inactive in presence of Mg2+, it shows 35% of maximal activity in presenceof Mn2+

D380A

site-directed mutagenesis, the mutant is inactive in presence of Mg2+

D380E

site-directed mutagenesis, the mutant is inactive in presence of Mg2+ or Mn2+

D531A

site-directed mutagenesis, the mutant is inactive in presence of Mg2+

D531E

site-directed mutagenesis, the mutant is inactive in presence of Mg2+, it shows 60% of maximal activity in presenceof Mn2+

R113A

Herpes simplex virus

mutation reduces viral yield (2fold), alters the kinetics of viral DNA replication, and decreases the fidelity of DNA replication

R182A

Herpes simplex virus

mutation reduces viral yield (2fold), alters the kinetics of viral DNA replication, and decreases the fidelity of DNA replication

R279A

Herpes simplex virus

mutation reduces viral yield (5fold), alters the kinetics of viral DNA replication, and decreases the fidelity of DNA replication

R280A

Herpes simplex virus

mutation reduces viral yield (5fold), alters the kinetics of viral DNA replication, and decreases the fidelity of DNA replication

A329A

not meaningfully associated with breast cancer risk; more likely to respond to Pt-based chemotherapy

A467T

naturally occuring mutation, the mutation is the most common POLG mutation and has been found to be associated with all of the disease symptoms analyzed. The A467T pol gamma possesses only 4% of the wild-type DNA polymerase activity and is compromised for its ability to interact with the p55 accessory subunit

A471V

moderate decrease in activity

A957S

naturally occuring mutation, involved in autosomal dominant progressive external ophthalmoplegia, the mutation is associated with motiif B in the active site

D115A/E116A

catalytically inactive

705693

D189G

impaired for extension step of TLS

D198A/E200A

site-directed mutagenesis in the exonuclease domain resulting in loss of exonuclease activity

D275A/E277A/D368A

Q07864; Q9NR33; P56282; Q9NRF9

mutation in the catalytic subunit, exonuclease-deficient variant

761518

E1143G

naturally occuring mutation, that is a frequent cause of ataxia-neuropathy syndrome, and found in 4% of European populations

E200A

exonuclease-deficient mutant

E292K

E29K

decreased insertion opposite abasic site (2-20)

E419G

20fold decrease in kcat/Km on dG and 670fold decrease on N2-CH2-Anth-dG, extension defect

E430G

low activity on AP site

E449K

low activity on AP site, low fidelity

F155S

decreased activity on model abasic site

F192C

F506G

inactive mutant enzyme

662621

F506R

complete loss of de novo DNA synthesis

662621

G154E

decreased activity opposite model abasic site, pathogenic

G848S

naturally occuring mutation, involved in Alpers syndrome, the mutant shows compromised DNA binding ability that affects its overall functional efficiency

G923D

naturally occuring mutation, involved in autosomal dominant progressive external ophthalmoplegia, the mutation is associated with motiif B in the active site

I39T

L21F

L442F

low activity on AP site

L606G

site-directed mutagenesis, the mutant shows increased polymerase activity and slightly reduced exonuclease activity compared to the wild-type enzyme, mutant pol delta L606G is highly error prone, incorporating single noncomplementary nucleotides at a high frequency during DNA synthesis

722686

L606K

site-directed mutagenesis, the mutant shows increased polymerase activity and reduced exonuclease activity compared to the wild-type enzyme, mutant pol delta L606K is extremely accurate, with a higher fidelity of single nucleotide incorporation by the active site than that of wild-type pol delta, it does not catalyze detectable nucleotide mis-insertion even with nucleotide concentrations as high as 4 mM, but pol delta L606K mutant is impaired in the bypass of DNA adducts

P169T

R219I

R246X

5-10fold less active with 8-oxo-dG-, N2-CH2-Anth-dG-, O6-Me-dG- and abasic-containing templates

R298H

R512W

decreased activity on undamaged and damaged DNA

R61A

Q9Y253

the mutations decreases the ability of the enzyme to distinguish between a thymine-thymine dimer lesion and undamaged DNA. Mutation does not impact the global structure and equilibrium motions of DNA polymerase eta. Only local conformational changes around the mutation sites are observed

R61K

R852C

naturally occuring mutation, involved in Alpers syndrome, the mutant shows compromised DNA binding ability that affects its overall functional efficiency

R853Q

naturally occuring mutation, involved in Alpers syndrome, the mutant shows compromised DNA binding ability that affects its overall functional efficiency

R943H

R964C

S423R

1.6fold more effcient than wild-type, 2fold increased DNA binding affinity, pathogenic

S62A

S62L

T44M

lesion-specific reduction in activity. Reduced activity with N2-CH2-Anth-dG, O6-Me-dG and abasic sites

T473A

decreased activity on undamaged and damaged DNA

T851A

naturally occuring mutation, involved in Alpers syndrome, the mutant shows compromised DNA binding ability that affects its overall functional efficiency

V130I

mutant has relaxed discrimination against the major groove adduct N6-furfuryl-dA

W748S

naturally occuring mutation that has intrinsic lower polymerase activity as well as a demonstrated lower affinity for DNA compared to the wild-type enzyme

W748S/E1143G

naturally occuring mutation, the E1143G single-nucleotide polymorphism can modulate the deleterious effect of the W748S mutation

Y271G

P06746

the mutant is rationally designed to provide flexibility to the steric gate backbone carboxyl of Tyr-271 in pol beta

Y432S

Y505A

Y955C

naturally occuring mutation, involved in autosomal dominant progressive external ophthalmoplegia, the mutation is associated with motiif B in the active site, the mutant retains less than 1% of the wild-type polymerase activity and displays a severe decrease in processivity, the mutation increases nucleotide misinsertion replication errors 10-100 fold in the absence of exonucleolytic proofreading

D907V

Human alphaherpesvirus 1

mutant is less resistant to acyclovir and cidofovir than the wild type enzyme

690480

L778M

Human alphaherpesvirus 1

mutant is less resistant to acyclovir and cidofovir than the wild type enzyme

690480

H329A

Mus musculus

mutation has little effect on template-dependent synthesis by Pol l from a paired primer terminus, but it reduces both template-independent and template-dependent synthesis during nonhomologous DNA end joining of intermediates whose 3' ends lack complementary template strand nucleotides

676140

D362A

Oryza sativa

the misincorporation frequency values of the D362A mutant are slightly higher (1.44.9fold) than those of the wild type protein

676558

A408S

A408S mutation results in a significant increase in both dNTP binding affinity and fidelity, kcat/Km for dATP is about 45% compared to the wild-type enzyme, D215A mutation results in inactivation of 3'-5'-exonuclease activity

D215A

3'-5'-exonuclease inactive mutant enzyme

D405A

mutant enzyme loses 99.8% of DNA polymerizing activity and 90% of 3'->5' exonucleolytic activity

D405E

mutant enzyme loses 95.8% of DNA polymerizing activity and 90% of 3'->5' exonucleolytic activity

D473G

wild-type variant Pfu-Pol(exo-) is is 60fold less accurate than Pfu-Pol(exo+)

662975

DELTAH672-S775

mutant enzyme loses 99% of DNA polymerizing activity and 97% of 3'->5' exonucleolytic activity

DELTAL717-S775

mutant enzyme loses 97% of DNA polymerizing activity and 97% of 3'->5' exonucleolytic activity

DELTAL746-S775

mutant protein has DNA polymerizing activity with 2.3fold higher specific activity than that of the wild-type but retains only 10% of the 3'->5' exonucleolytic activity of the wild-type

L409I

kcat/Km for dATP is about 20% compared to the wild-type enzyme

L409M

L409 mutation results in drastically reduced affinity for the correct dNTP, a much higher efficiency of both misincorporation and mismatch extension, and substantially lower fidelity as demonstrated by a PCR-based forward mutation assay, kcat/Km for dATP is about 155% compared to the wild-type enzyme, D215A mutation results in inactivation of 3'-5'-exonuclease activity

L409V

kcat/Km for dATP is about 35% compared to the wild-type enzyme

T471A

less accurate, by factors of 1.6 than the wild-type variant Pfu-Pol(exo-)

662975

T471G

less accurate, by factors of 1.2 than the wild-type variant Pfu-Pol(exo-)

662975

Y410I

kcat/Km for dATP is about 20% compared to the wild-type enzyme

Y410L

kcat/Km for dATP is about 45% compared to the wild-type enzyme

Y410V

Y410V mutation results in high fidelity in both misincorporation assays and forward mutation assays, but displays a substantially higher Km than wild-type enzyme, kcat/Km for dATP is about 10% compared to the wild-type enzyme, D215A mutation results in inactivation of 3'-5'-exonuclease activity

D1122A

loss of polymerization activity

D1122E

mutant enzyme with reduced polymerization activity, polymerization activity is 15% compared to wild-type activity, 3'-5' exonuclease activity remains, the mutant has lower Mg2+ affinity than does the wild-type

D1124A

loss of polymerization activity

D1124E

mutant enzyme with reduced polymerization activity, polymerization activity is 43% compared to wild-type activity,

D1124N

mutant enzyme with reduced polymerization activity, 3'-5' exonuclease activity remains, the mutant has lower Mg2+ affinity than does the wild-type

D259E

moderate decrease of the polymerizing activity

D259G

moderate decrease of the polymerizing activity

D259K

the exonuclease activity of the mutant enzyme decreases drastically to 0.58% compared with that of the wild-type DNA polymerase

D259N

moderate decrease of the polymerizing activity

G1123A

mutant enzyme has 13% of the activity of the wild type enzyme

G1123R

mutant enzyme has 0.8% of the activity of the wild type enzyme

K253E

exonuclease activity of mutant increases 2.7fold compared to wild-type activity

K253E/R255E

exonuclease activity of mutant increases 1.8fold compared to wild-type activity

R255D

exonuclease activity of mutant increases 2.9fold compared to wild-type activity

R258A |

Rattus norvegicus

mutant of Pol beta has a facilitated subdomain-reopening step. Rate of single-nucleotide incorporation catalyzed by R258A is identical to that of wild-type

672254

D424A

P95979

mutation in polymerase active site, the polymerase activity is reduced more than 30fold compared to the wild-type activity

718921

D424A/D542A

P95979

mutation in polymerase active site, complete inactivation of polymerase activity

718921

D542A

P95979

mutation in polymerase active site, the polymerase activity is reduced more than 50fold compared to the wild-type activity

718921

F12A

3 entries

N188W

kcat/Km for dCTP is 2.9fold compared to kcat/Km of wild-type enzyme

725385

N249Y

exhibits increased catalytic activity when compared to the wild-type enzyme, the mutation decreases the affinity for NAD(H) cofactor

725713

R322H

the His332 mutant exhibits faster forward rate constants relative to wild-type Dpo4. The kpol values for the His332 mutant incorporation opposite G and 8-oxoG are 3.6- and 4.6fold faster than for wild-type Dpo4. The nucleotide binding affinity trend is opposite that of wild-type Dpo4, Glu332, and Leu332, with tighter dCTP binding during bypass of G. As in the case of Ala332, the kinetic analysis indicates that His332 inserts dCTP opposite G with slightly greater efficiency than opposite 8-oxoG

R322L

kpol (forward rate of polymerization) values for the Leu332 mutant incorporation opposite G and 8-oxoG are 4.1- and 1.9fold higher than those for wild-type Dpo4. The Leu332 mutant is about 2fold more efficient at incorporation of dCTP opposite 8-oxoG compared with G

R331A/R322A

mutant has lower forward rate constants relative to wild-type enzyme for both G and 8-oxoG. The double mutant-catalyzed insertion of dCTP opposite 8-oxoG is about 4fold higher than dCTP insertion opposite G. The measured binding affinity of dCTP is tighter than that of wild-type Dpo4 for unmodified DNA, but the binding affinity of dCTP opposite 8-oxoG is similar to that observed for wild-type enzyme. The catalytic efficiency for dCTP incorporation increases about 4fold for unmodified DNA and decreases about 2fold for 8-oxoG-modified DNA. The mutant fails to incorporate dATP opposite 8-oxoG in the presteady-state experiments. It inserts dCTP opposite 8-oxoG with an about 200fold greater efficiency than it does dATP and the steady-state efficiency of dATP incorporation is decreased about 27fold relative to the wild-type enzyme

R332A

mutant enzyme displays a higher affinity (lower KD,dCTP) for dCTP when bound to the unmodified DNA compared with the KD,dCTP measured for mutant-catalyzed incorporation of dCTP opposite 8-oxoG. Wild-type enzyme shows a greater affinity for dCTP opposite to 8-oxoG-modified DNA. The catalytic efficiency of the mutant is 23fold higher at incorporation of C opposite G and 1.2fold lower than wild-type enzyme in dCTP incorporation opposite 8-oxoG

R332E

mutant enzyme retains fidelity against bypass of 7,8-dihydro-8-oxodeoxyguanosine (8-oxoG) that is similar to wild enzyme. A crystal structure of the mutant and 8-oxoG:C pair reveals water-mediated hydrogen bonds between Glu332 and the O-8 atom of 8-oxoG. The kpol (forward rate of polymerization) value for dCTP incorporation opposite G is 4.8fold higher than for wild-type enzyme. The kpol (forward rates of polymerization) value for dCTP incorporation opposite 8-oxoG is 3.5fold higher than wild-type enzyme insertion of dCTP opposite 8-oxoG. The catalytic efficiency of the Glu332 mutant is 2.3fold greater than wild-type Dpo4 for dCTP incorporation opposite G but 1.7fold less efficient than wild-type Dpo4 for incorporation opposite 8-oxoG

T239W

3 entries

Y12A

Saccharolobus solfataricus P1

F12A

no detectable incorporation of ribonucleotide triphosphate is observed with the wild-type enzyme, the mutant form F12A efficiently incorporates and extends ribonucleotide triphosphate, even in the absence of Mn2+ ions

D424A

mutation in polymerase active site, the polymerase activity is reduced more than 30fold compared to the wild-type activity

D424A/D542A

mutation in polymerase active site, complete inactivation of polymerase activity

D542A

mutation in polymerase active site, the polymerase activity is reduced more than 50fold compared to the wild-type activity

F12A

N188W

kcat/Km for dCTP is 2.9fold compared to kcat/Km of wild-type enzyme

N249Y

exhibits increased catalytic activity when compared to the wild-type enzyme, the mutation decreases the affinity for NAD(H) cofactor

R322H

R322L

R331A/R322A

R332A

R332E

T239W

3 entries

Y12A

M644F

mutant enzyme has reduced fidelity resulting from strongly increased misinsertion rates

676222

M644L

mutant enzyme synthesizes DNA with high fidelity

676222

M644W

mutant enzyme synthesizes DNA with high fidelity

676222

Y708A

mutation of pol delta, exhibits slow growth, sensitivity to hydroxyurea and strong mutator phenotype for frameshifts and base substitutions

643664

Y831A

mutation of pol epsilon, slight sensitivity to hydroxyurea, semidominant mutator phenotype for frameshifts and base substitutions

643664

Y869A

mutation of pol alpha, strain is viable, exhibits slow growth, sensitivity to hydroxyurea and spontaneous mutator phenotype for frameshifts and base substitutions

643664

I364Q

Salasvirus phi29

binds the substrate with less efficiency than wild-type enzyme

643672

I364R

Salasvirus phi29

unable binding of the substrate to the enzyme

643672

K366T

Salasvirus phi29

mutant enzyme shows a wild-type like phenotype in DNA-primed polymerisation in the presence of DNA as template, in terminal protein-primed reactions as initiation and transition it is impaired, especially in the presence of the Phi29 DNA-binding protein, protein p6

662613

K371T

Salasvirus phi29

binds the substrate with the same efficiency as wild-type enzyme

643672

D10A

Streptococcus pneumoniae

retains polymerase activity, reduced exonuclease activity, changes in dependency on metal activation of exonuclease activity

643653

D190A

Streptococcus pneumoniae

retains polymerase activity, reduced exonuclease activity, changes in dependency on metal activation of exonuclease activity

643652

K242R/I243K/P244S

D111A

94% loss of DNA polymerase activity

D171A

96% loss of DNA polymerase activity

E113A

92% loss of DNA polymerase activity

E128A

23% loss of DNA polymerase activity

H145A

98% loss of DNA polymerase activity

H190A

31% loss of DNA polymerase activity

K65A

98% loss of DNA polymerase activity

K66A

70% loss of DNA polymerase activity

K70A

19% loss of DNA polymerase activity

N176A

2% increase of DNA polymerase activity

S184A

46% loss of DNA polymerase activity

T134A

23% loss of DNA polymerase activity

T139A

83% loss of DNA polymerase activity

Y147A

19% loss of DNA polymerase activity

Y178A

40% loss of DNA polymerase activity

Y46A

2% loss of DNA polymerase activity

Thermococcus kodakarensis

D112A/E114A

Tequatrovirus T4

the mutant T4 DNA polymerases is defective in 3'-5' exonuclease activity and has a 1000fold increase in the development of spontaneous mutations compared to wild type polymerase

703192

D219A

Tequatrovirus T4

the mutant T4 DNA polymerases is defective in 3'-5' exonuclease activity and has a 1000fold increase in the development of spontaneous mutations compared to wild type polymerase

703192

D324A

Tequatrovirus T4

the mutant T4 DNA polymerases is defective in 3'-5' exonuclease activity and has a 1000fold increase in the development of spontaneous mutations compared to wild type polymerase, the exonuclease activity of the D324A mutant is 100000-fold lower than wild type polymerase

703192

N210D

P77933

the mutant enzyme shows very weak 3'-5'-exonuclease activity compared to the wild-type enzyme (0.1%). No significant difference in DNA polymerase activity as compared with that of the wild-type enzyme. Mutation frequency in PCR becomes higher as 3'5' exonuclease activity decreases

725590

Y311F

Thermococcus kodakarensis

P77933

the mutant enzyme shows very weak 3'-5'-exonuclease activity compared to the wild-type enzyme (0.01%). No significant difference in DNA polymerase activity as compared with that of the wild-type enzyme

725590

molecular biology

Thermococcus waiotapuensis

H633D

H9CW54

DNA polymerase activity of the mutant enzyme is higher than the polymerase activity of the wild-type enzyme. PCR efficiency of the H633D mutant is higher than that of the N565K mutant enzyme

729659

H633R

Thermococcus waiotapuensis

H9CW54

significantly improved polymerase function compared to wild-type enzyme in terms of processivity (2-fold), extension rate (1.5fold) and PCR efficiency. The kcat value of the Twa H633R mutant is similar to that of wild-type, but the Km of the Twa H633R mutant is about 1.6fold lower than that of the wild-type

729659

N565K

Thermococcus waiotapuensis

H9CW54

DNA polymerase activity of the mutant enzyme is higher than the polymerase activity of the wild-type enzyme. PCR efficiency of the H633D mutant is higher than that of the N565K mutant enzyme

729659

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

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

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

E602V/A608V/I614M/E615G

P19821

the mutant enzyme is able to incorporate both NTPs and dNTPs with the same catalytic efficiency as the wild-type enzyme incorporates dNTPs. The mutant enzyme allowed the generation of mixed RNADNA amplification products in PCR demonstrating DNA polymerase, RNA polymerase as well as reverse transcriptase activity within the same polypeptide. The mutant displays an expanded substrate spectrum towards other 2'-substituted nucleotides and is able to synthesize nucleic acid polymers in which each base bear a different 2'-substituent

675398

G418K

increased exonuclease activity

643660

L831N/A814R

P19821

truncated enzyme delta413-L813N/A814R has reduced temperature stability

643675

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 2.7.7.49

723163

Q507E