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Literature summary extracted from

  • Choi, J.Y.; Eoff, R.L.; Pence, M.G.; Wang, J.; Martin, M.V.; Kim, E.J.; Folkmann, L.M.; Guengerich, F.P.
    Roles of the four DNA polymerases of the crenarchaeon Sulfolobus solfataricus and accessory proteins in DNA replication (2011), J. Biol. Chem., 286, 31180-31193.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.7.7 proliferating cell nuclear antigen DNA synthesis by any of the DNA polymerases alone is not very processive. The addition of proliferating cell nuclear antigen slightly increases primer extension only by DNA polymerase Dpo4 but not by DNA polymerase Dpo1, Dpo2, or Dpo3. Both proliferating cell nuclear antigen and replication factor C enhance primer extension substantially in the cases of DNA polymerase Dpo1, Dpo3, and Dpo4 (which generate much longer extension products up to about 150-, 65-, and 150-mers, respectively) but very weakly with DNA polymerase Dpo2, which generates slightly more products of similar length up to about 80-mers. The addition of proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein strongly increases DNA polymerization by Dpo1 and Dpo4, which yields extended products, mainly up to about 500-mers and 300-mers, respectively. An increase of the processivity of DNA synthesis in the presence of all accessory replication factors is the most prominent with Dpo1 and also with Dpo4 Saccharolobus solfataricus
2.7.7.7 Replication factor C both proliferating cell nuclear antigen and replication factor C enhance primer extension substantially in the cases of DNA polymerase Dpo1, Dpo3, and Dpo4 (which generate much longer extension products up to about 150-, 65-, and 150-mers, respectively) but very weakly with DNA polymerase Dpo2, which generates slightly more products of similar length up to about 80-mers. The addition of proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein strongly increases DNA polymerization by Dpo1 and Dpo4, which yields extended products, mainly up to about 500-mers and 300-mers, respectively. An increase of the processivity of DNA synthesis in the presence of all accessory replication factors (proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein) is the most prominent with Dpo1 and also with Dpo4 Saccharolobus solfataricus
2.7.7.7 single-stranded binding protein an increase of the processivity of DNA synthesis in the presence of all accessory replication factors (proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein) is the most prominent with Dpo1 and also with Dpo4 as compared with DNA polymerase Dpo2 and Dpo3 Saccharolobus solfataricus
2.7.7.7 single-stranded binding protein an increase of the processivity of DNA synthesis in the presence of all accessory replication factors (proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein) is the most prominent with Dpo1 and also with Dpo4 as compared woth DNA polymerase Dpo2 and Dpo3 Saccharolobus solfataricus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7 expression in Escherichia coli Saccharolobus solfataricus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.7 Replication factor C DNA polymerization by Dpo2 and Dpo3 is strongly inhibited; DNA polymerization by Dpo2 and Dpo3 is strongly inhibited Saccharolobus solfataricus
2.7.7.7 single-stranded binding protein DNA polymerization by Dpo2 and Dpo3 is strongly inhibited; DNA polymerization by Dpo2 and Dpo3 is strongly inhibited; DNA synthesis by DNA polymerase Dpo2 and Dpo3 is remarkably decreased by single-stranded binding protein; DNA synthesis by DNA polymerase Dpo2 and Dpo3 is remarkably decreased by single-stranded binding protein Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.7 0.0084
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.0098
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo2 Saccharolobus solfataricus
2.7.7.7 0.039
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.075
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.19
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.39
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.47
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.69
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.93
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 1.6
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 2.1
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 2.5
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 3.1
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 3.7
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo2 Saccharolobus solfataricus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.7 Mg2+ DNA polymerase Dpo2 and Dpo3 are both more active with Mg2+ than Mn2+. DNA polymerase Dpo1 and Dpo4 are similarly active with Mg2+ or Mn2+ Saccharolobus solfataricus
2.7.7.7 Mn2+ DNA polymerase Dpo2 and Dpo3 are both more active with Mg2+ than Mn2+. DNA polymerase Dpo1 and Dpo4 are similarly active with Mg2+ or Mn2+ Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Saccharolobus solfataricus P26811
-
-
2.7.7.7 Saccharolobus solfataricus P95979
-
-
2.7.7.7 Saccharolobus solfataricus Q07635
-
-
2.7.7.7 Saccharolobus solfataricus Q97W02
-
-
2.7.7.7 Saccharolobus solfataricus P2 P26811
-
-
2.7.7.7 Saccharolobus solfataricus P2 P95979
-
-
2.7.7.7 Saccharolobus solfataricus P2 Q07635
-
-
2.7.7.7 Saccharolobus solfataricus P2 Q97W02
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.7
-
 Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 dATP + DNAn
-
 Saccharolobus solfataricus ?
-
 ?
2.7.7.7 dATP + DNAn
-
 Saccharolobus solfataricus P2 ?
-
 ?
2.7.7.7 dCTP + DNAn
-
 Saccharolobus solfataricus ?
-
 ?
2.7.7.7 dCTP + DNAn
-
 Saccharolobus solfataricus P2 ?
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn proliferating cell nuclear antigen facilitates DNA synthesis with Dpo3, as with Dpo1 and Dpo4, but very weakly with Dpo2. DNA synthesis in the presence of proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein is most processive with DNA polymerase Dpo1 in comparison to DNA polymerase Dpo3 and Dpo4. DNA lesion bypass DNA synthesis in the presence of proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein is most effective with DNA polymerase Dpo4 in comparison to DNA polymerase Dpo1 and Dpo3. Both Dpo2 and Dpo3, but not Dpo1, bypass hypoxanthine and 8-oxoguanine. Dpo2 and Dpo3 bypass uracil and cis-syn cyclobutane thymine dimer, respectively. DNA polymerase Dpo2 and Dpo3 possess very low DNA polymerase and 3' to 5' exonuclease activities in vitro compared with Dpo1 and Dpo4 Saccharolobus solfataricus diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn proliferating cell nuclear antigen facilitates DNA synthesis with Dpo3, as with Dpo1 and Dpo4, but very weakly with Dpo2. DNA synthesis in the presence of proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein is most processive with DNA polymerase Dpo1 in comparison to DNA polymerase Dpo3 and Dpo4. DNA lesion bypass DNA synthesis in the presence of proliferating cell nuclear antigen, replication factor C, and single-stranded binding protein is most effective with DNA polymerase Dpo4 in comparison to DNA polymerase Dpo1 and Dpo3. Both Dpo2 and Dpo3, but not Dpo1, bypass hypoxanthine and 8-oxoguanine. Dpo2 and Dpo3 bypass uracil and cis-syn cyclobutane thymine dimer, respectively. DNA polymerase Dpo2 and Dpo3 possess very low DNA polymerase and 3' to 5' exonuclease activities in vitro compared with Dpo1 and Dpo4 Saccharolobus solfataricus P2 diphosphate + DNAn+1
-
 ?
2.7.7.7 dGTP + DNAn
-
 Saccharolobus solfataricus ?
-
 ?
2.7.7.7 dGTP + DNAn
-
 Saccharolobus solfataricus P2 ?
-
 ?
2.7.7.7 dTTP + DNAn
-
 Saccharolobus solfataricus ?
-
 ?
2.7.7.7 dTTP + DNAn
-
 Saccharolobus solfataricus P2 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.7 DNA polymerase
-
 Saccharolobus solfataricus
2.7.7.7 Dpo1
-
 Saccharolobus solfataricus
2.7.7.7 Dpo2
-
 Saccharolobus solfataricus
2.7.7.7 Dpo3
-
 Saccharolobus solfataricus
2.7.7.7 Dpo4
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.7 37
-
 assay at Saccharolobus solfataricus
2.7.7.7 50
-
-
 Saccharolobus solfataricus
2.7.7.7 60
-
-
 Saccharolobus solfataricus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.7 25 60 the catalytic activities of DNA polymerase Dpo2 gradually increases from 25 to 50°C but abruptly decreases at 60°C, at which it is only20% of the activity at 50°C (and even lower than the activities at 37°C) and is abolished at 70 °C Saccharolobus solfataricus
2.7.7.7 25 60 the catalytic activity of DNA polymerase Dpo3 gradually increases from 25 to 50°C but abruptly decreases at 60°C, at which it is 20% of the activity at 50°C (and even lower than the activity at 37°C) and is abolished at 70°C Saccharolobus solfataricus
2.7.7.7 25 70 activities of DNA polymerase Dpo1 gradually increases from 25 to 60°C and is substantial even at 70°C, with40% of the activity at 60°C Saccharolobus solfataricus
2.7.7.7 25 70 activity of DNA polymerase Dpo4 gradually increases from 25 to 60°C and is substantial even at 70°C, with60% of the activity at 60°C Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.7 additional information
-
 DNA polymerase Dpo2 and Dpo3 are more thermolabile than Dpo1 and Dpo4 Saccharolobus solfataricus
2.7.7.7 60
-
 50 mM potassium HEPES buffer (pH 7.5) without any stabilizing agents. Activities of DNA polymerase Dpo2 and Dpo3 abruptly decrease to 5% of control activity following heat preincubation for only 5 and 10 min, respectively, and are completely abolished after 20 min of heat preincubation. Activities of DNA polymerase Dpo1 and Dpo4 are86% and 20% of control activity, respectively, even after 40 min of heat preincubation Saccharolobus solfataricus
2.7.7.7 60
-
 50 mM potassium HEPES buffer (pH 7.5) without any stabilizing agents. Activity of DNA polymerase Dpo1 is86% of control activity, respectively, even after 40 min of heat preincubation Saccharolobus solfataricus
2.7.7.7 60
-
 50 mM potassium HEPES buffer (pH 7.5) without any stabilizing agents. Activity of DNA polymerase Dpo3 abruptly decreases to 5% of control activity following heat preincubation for only 5 and 10 min, respectively, and is completely abolished after 20 min of heat preincubation Saccharolobus solfataricus
2.7.7.7 60
-
 50 mM potassium HEPES buffer (pH 7.5) without any stabilizing agents. Activity of DNA polymerase Dpo4 is 20% of control activity even after 40 min of heat preincubation Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.7 0.0000072
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo2 Saccharolobus solfataricus
2.7.7.7 0.000013
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.000016
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.000053
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo2 Saccharolobus solfataricus
2.7.7.7 0.00011
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.00053
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.0018
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.0068
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.073
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.096
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.15
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.23
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.48
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 12
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.7 7.5
-
 assay at Saccharolobus solfataricus

Expression

EC Number Organism Comment Expression
2.7.7.7 Saccharolobus solfataricus constitutively expressed in all growth phases. The expression levels of DNA polymerase Dpo1 and Dpo3 are an order of magnitude higher than for Dpo2 and Dpo4 additional information

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.7.7 0.0000000019
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo2 Saccharolobus solfataricus
2.7.7.7 0.000000041
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.000000068
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.00000012
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.00000086
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.0000027
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.0000054
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo2 Saccharolobus solfataricus
2.7.7.7 0.0000071
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo3 Saccharolobus solfataricus
2.7.7.7 0.000031
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.00011
-
 dATP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.0003
-
 dTTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.00032
-
 dGTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus
2.7.7.7 0.027
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo1 Saccharolobus solfataricus
2.7.7.7 0.31
-
 dCTP pH 7.5, 37°C, DNA polymerase Dpo4 Saccharolobus solfataricus