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Literature summary extracted from

  • Yasukawa, K.; Konishi, A.; Shinomura, M.; Nagaoka, E.; Fujiwara, S.
    Kinetic analysis of reverse transcriptase activity of bacterial family A DNA polymerases (2012), Biochem. Biophys. Res. Commun., 427, 654-658.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7 expression of soluble mutant L329A in Escherichia coli strain BL21(DE3) codon plus Thermotoga petrophila
2.7.7.7 N-terminally His6-tagged M1 DNA polymerase in Eschrichia coli strain BL21(DE3) codon plus Thermus thermophilus
2.7.7.49 expression of the C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Moloney murine leukemia virus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.7 L329A site-directed mutagenesis Thermotoga petrophila

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.7 additional information high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases Thermotoga petrophila
2.7.7.7 additional information high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases Thermus thermophilus
2.7.7.7 poly(rA)-p(dT)45 inhibitory effect towards the reverse transcriptase activity of K4polL329A Thermotoga petrophila
2.7.7.7 poly(rA)-p(dT)45 inhibitory effect towards the reverse transcriptase activity of M1pol Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.7 additional information
-
 additional information the reaction rate of K4polL329A exhibits a saturated profile of the Michaelis-Menten kinetics for dTTP concentrations but a substrate inhibition profile for poly(rA)-p(dT)45 concentrations Thermotoga petrophila
2.7.7.7 additional information
-
 additional information the reaction rate of M1pol exhibits a saturated profile of the Michaelis-Menten kinetics for dTTP concentrations but a substrate inhibition profile for poly(rA)-p(dT)45 concentrations Thermus thermophilus
2.7.7.7 0.23
-
 dTTP pH 8.2, 50°C, recombinant M1pol Thermus thermophilus
2.7.7.7 0.28
-
 dTTP pH 8.2, 50°C, recombinant enzyme mutant K4polL329A Thermotoga petrophila
2.7.7.49 additional information
-
 additional information Michaelis-Menten kinetics Moloney murine leukemia virus
2.7.7.49 0.0056
-
 DNAn pH 8.2, 37°C, recombinant enzyme, substrate is poly(rA)-p(dT)45 Moloney murine leukemia virus
2.7.7.49 0.25
-
 dTTP pH 8.2, 37°C, recombinant enzyme Moloney murine leukemia virus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn Thermotoga petrophila
-
 diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn Thermus thermophilus
-
 diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn Thermus thermophilus M1
-
 diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn Thermotoga petrophila K4
-
 diphosphate + DNAn+1
-
 ?
2.7.7.7 additional information Thermotoga petrophila only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 ?
-
 ?
2.7.7.7 additional information Thermotoga petrophila K4 only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 ?
-
 ?
2.7.7.49 deoxynucleoside triphosphate + DNAn Moloney murine leukemia virus
-
 diphosphate + DNAn+1
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Thermotoga petrophila
-
-
-
2.7.7.7 Thermotoga petrophila K4
-
-
-
2.7.7.7 Thermus thermophilus K4Q1U9 isolated from a hot spring at Kagoshima, Japan
-
2.7.7.7 Thermus thermophilus M1 K4Q1U9 isolated from a hot spring at Kagoshima, Japan
-
2.7.7.49 Moloney murine leukemia virus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.7 N-terminally His6-tagged M1 DNA polymerase from Eschrichia coli strain BL21(DE3) codon plus by nickel affinity chromatography and dialysis Thermus thermophilus
2.7.7.7 recombinant soluble mutant L329A from Escherichia coli strain BL21(DE3) codon plus partially by heat treatment at 85°C for 30 min Thermotoga petrophila

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermotoga petrophila diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermus thermophilus diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn DNA-dependent DNA polymerase activity to incorporate dNTP into gapped M13mp2 DNA Thermus thermophilus diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermus thermophilus M1 diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn DNA-dependent DNA polymerase activity to incorporate dNTP into gapped M13mp2 DNA Thermus thermophilus M1 diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermotoga petrophila K4 diphosphate + DNAn+1
-
 ?
2.7.7.7 dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermotoga petrophila diphosphate + DNAn+1
-
 ?
2.7.7.7 dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermus thermophilus diphosphate + DNAn+1
-
 ?
2.7.7.7 dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermus thermophilus M1 diphosphate + DNAn+1
-
 ?
2.7.7.7 dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermotoga petrophila K4 diphosphate + DNAn+1
-
 ?
2.7.7.7 additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 Thermotoga petrophila ?
-
 ?
2.7.7.7 additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, no activity with the wild-type enzyme Thermotoga petrophila ?
-
 ?
2.7.7.7 additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 Thermotoga petrophila K4 ?
-
 ?
2.7.7.7 additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, no activity with the wild-type enzyme Thermotoga petrophila K4 ?
-
 ?
2.7.7.49 deoxynucleoside triphosphate + DNAn
-
 Moloney murine leukemia virus diphosphate + DNAn+1
-
 ?
2.7.7.49 dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Moloney murine leukemia virus diphosphate + DNAn+1
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.7 K4pol
-
 Thermotoga petrophila
2.7.7.7 M1 DNA polymerase
-
 Thermus thermophilus
2.7.7.7 M1pol
-
 Thermus thermophilus
2.7.7.49 MMLV RT
-
 Moloney murine leukemia virus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.7 50
-
 assay at Thermotoga petrophila
2.7.7.7 50
-
 assay at Thermus thermophilus
2.7.7.49 37
-
 assay at Moloney murine leukemia virus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.7 37 65 optimum above Thermotoga petrophila

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.7 1
-
 dTTP pH 8.2, 50°C, recombinant M1pol Thermus thermophilus
2.7.7.7 1.7
-
 dTTP pH 8.2, 50°C, recombinant enzyme mutant K4polL329A Thermotoga petrophila
2.7.7.49 29
-
 DNAn pH 8.2, 37°C, recombinant enzyme, substrate is poly(rA)-p(dT)45 Moloney murine leukemia virus
2.7.7.49 35
-
 dTTP pH 8.2, 37°C, recombinant enzyme Moloney murine leukemia virus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.7 8.2
-
 assay at Thermotoga petrophila
2.7.7.7 8.2
-
 assay at Thermus thermophilus
2.7.7.49 8.2
-
 assay at Moloney murine leukemia virus

General Information

EC Number General Information Comment Organism
2.7.7.7 evolution the enzyme belongs to the DNA polymerase family A Thermotoga petrophila
2.7.7.7 evolution the enzyme belongs to the DNA polymerase family A Thermus thermophilus
2.7.7.7 additional information nine residues, Tyr326, Leu329, Gln384, Lys387, Phe388, Met408, Asn422, Tyr438, and Phe451, are predicted to be involved in DNA/RNA distinction Thermotoga petrophila