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Literature summary extracted from

  • Prato, S.; Vitale, R.M.; Contursi, P.; Lipps, G.; Saviano, M.; Rossi, M.; Bartolucci, S.
    Molecular modeling and functional characterization of the monomeric primase-polymerase domain from the Sulfolobus solfataricus plasmid pIT3 (2008), FEBS J., 275, 4389-4402.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.B16 purified following overexpression in Escherichia coli Saccharolobus solfataricus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.B16 Mg2+ the enzyme is strictly dependent on divalent cations, activating metal preferably used by Rep245 for its DNA polymerase activity is Mg2+ at concentrations between 5 and 10 mM Saccharolobus solfataricus
2.7.7.B16 Mn2+ the enzyme is strictly dependent on divalent cations, with Mn2+ as a cofactor, the DNA polymerase activity of Rep245 is optimal at 1–2.5 mM and decreases noticeably at increasing amounts of Mn2+ Saccharolobus solfataricus
2.7.7.B16 additional information Rep245 polymerase activity is strictly dependent on divalent cations (Mg2+ or Mn2+), Zn2+ cations do not support the DNA polymerization activity of Rep245 Saccharolobus solfataricus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.B16 29000
-
 x * 29000, Rep245 domain containing the N-terminal domain of the pIT3 replication protein encompassing residues 31–245, SDS-PAGE Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.B16 NTP + n NTP Saccharolobus solfataricus
-
 N(pN)n + n diphosphate
-
 ?
2.7.7.B16 NTP + n NTP Saccharolobus solfataricus P2
-
 N(pN)n + n diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.B16 Saccharolobus solfataricus Q9UWW1 and Q97Z83 Q9UWW1: large subunit, Q97Z83: small subunit
-
2.7.7.B16 Saccharolobus solfataricus P2 Q9UWW1 and Q97Z83 Q9UWW1: large subunit, Q97Z83: small subunit
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.B16 purified following overexpression in Escherichia coli Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.B16 NTP + n NTP
-
 Saccharolobus solfataricus N(pN)n + n diphosphate
-
 ?
2.7.7.B16 NTP + n NTP de novo synthesis of larger molecular size RNA products when incubated with M13 mp18 single-stranded DNA in the presence of a ribonucleotide mixture. The Rep245 domain is capable of utilizing both ribonucleotides and deoxyribonucleotides for de novo primer synthesis and it synthesizes DNA products up to several kb in length in a template-dependent manner. The Rep245 primase-polymerase domain harbors also a terminal nucleotidyl transferase activity, being able to elongate the 3'-end of synthetic oligonucleotides in a non-templated manner. The Rep245 domain contains the catalytic residues required for both primase and polymerase activities Saccharolobus solfataricus N(pN)n + n diphosphate
-
 ?
2.7.7.B16 NTP + n NTP
-
 Saccharolobus solfataricus P2 N(pN)n + n diphosphate
-
 ?
2.7.7.B16 NTP + n NTP de novo synthesis of larger molecular size RNA products when incubated with M13 mp18 single-stranded DNA in the presence of a ribonucleotide mixture. The Rep245 domain is capable of utilizing both ribonucleotides and deoxyribonucleotides for de novo primer synthesis and it synthesizes DNA products up to several kb in length in a template-dependent manner. The Rep245 primase-polymerase domain harbors also a terminal nucleotidyl transferase activity, being able to elongate the 3'-end of synthetic oligonucleotides in a non-templated manner. The Rep245 domain contains the catalytic residues required for both primase and polymerase activities Saccharolobus solfataricus P2 N(pN)n + n diphosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.7.B16 ? x * 29000, Rep245 domain containing the N-terminal domain of the pIT3 replication protein encompassing residues 31–245, SDS-PAGE Saccharolobus solfataricus

Synonyms

EC Number Synonyms Comment Organism
2.7.7.B16 pIT3 replication protein the Rep245 protein is the N-terminal domain of the pIT3 replication protein encompassing residues 31–245 Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.B16 65
-
-
 Saccharolobus solfataricus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.B16 45 75 45°C: about 40% of maximal activity, 75°C: about 40% of maximal activity Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.B16 80
-
 15 min, 40% loss of activity Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.B16 8
-
 maximal DNA template elongation at pH 8.0 Saccharolobus solfataricus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.B16 7 8.5 pH 7.0: about 40% of maximal activity, pH 8.5: about 50% of maximal activity Saccharolobus solfataricus