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Literature summary for 2.7.7.B16 extracted from

  • Lao-Sirieix, S.H.; Bell, S.D.
    The heterodimeric primase of the hyperthermophilic archaeon Sulfolobus solfataricus possesses DNA and RNA primase, polymerase and 3-terminal nucleotidyl transferase activities (2004), J. Mol. Biol., 344, 1251-1263.
    View publication on PubMed
Search for more literature for 2.7.7.B16

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
D101A/D103A the catalyric site mutant enzyme does not exhibit any detectable catalytic activity, even when higher concentrations of enzyme are utilised. The result demonstrate that the terminal transferase-like activity of PriSL is dependent on the catalytic activity of the primase. D101 and D103 are necessary for PriSL catalytic activity Saccharolobus solfataricus
N175A/R176 the affinity of DNA-binding site mutant for NTPs is approximately tenfold lower than that of the wild-type primase and that its enzymatic capability is diminished. Therefore, the mutation of N175 and R176 does not alter the DNA binding properties of the primase but modifies its affinity for free NTPs Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.025
-
 NTP pH 8.0, 60°C, in presence of M13 ssDNA as template Saccharolobus solfataricus
0.028
-
 dNTP pH 8.0, 60°C, in presence of M13 ssDNA as template Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ the enzyme requires manganese ions for catalytic activity Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus Q9UWW1 and Q97Z83 Q9UWW1: large subunit, Q97Z83: small subunit
-
Saccharolobus solfataricus P2 Q9UWW1 and Q97Z83 Q9UWW1: large subunit, Q97Z83: small subunit
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dNTP + n dNTP the oligonucleotides 20-mer oligo(dA), -(dC), -(dG) and -(dT) are used with varying efficiencies, with oligo(d)A being the poorest substrate and oligo(dC) the most preferred one. The enzyme is capable of utilising both ribonucleotides and deoxyribonucleotides for primer synthesis in the presence of natural, or synthetic, single-stranded DNA. It has a significantly higher affinity for ribonucleotides than for deoxyribonucleotides. In addition to the primase and polymerase activities, the enzyme possesses a template-independent 3'-terminal nucleotidyl transferase activity Saccharolobus solfataricus dN(pdN)n + n diphosphate
-
 ?
dNTP + n dNTP the oligonucleotides 20-mer oligo(dA), -(dC), -(dG) and -(dT) are used with varying efficiencies, with oligo(d)A being the poorest substrate and oligo(dC) the most preferred one. The enzyme is capable of utilising both ribonucleotides and deoxyribonucleotides for primer synthesis in the presence of natural, or synthetic, single-stranded DNA. It has a significantly higher affinity for ribonucleotides than for deoxyribonucleotides. In addition to the primase and polymerase activities, the enzyme possesses a template-independent 3'-terminal nucleotidyl transferase activity Saccharolobus solfataricus P2 dN(pdN)n + n diphosphate
-
 ?
NTP + n NTP
-
 Saccharolobus solfataricus N(pN)n + n diphosphate
-
 ?
NTP + n NTP the oligonucleotides 20-mer oligo(dA), -(dC), -(dG) and -(dT) are used as template with varying efficiencies, with oligo(d)A being the poorest substrate and oligo(dC) the most preferred one. The enzyme is capable of utilising both ribonucleotides and deoxyribonucleotides for primer synthesis in the presence of natural, or synthetic, single-stranded DNA. It has a significantly higher affinity for ribonucleotides than for deoxyribonucleotides. Products with an apparent length of 14 nt and 7 nt are mainly synthesised. In addition to the primase and polymerase activities, the enzyme possesses a template-independent 3'-terminal nucleotidyl transferase activity Saccharolobus solfataricus N(pN)n + n diphosphate
-
 ?
NTP + n NTP
-
 Saccharolobus solfataricus P2 N(pN)n + n diphosphate
-
 ?
NTP + n NTP the oligonucleotides 20-mer oligo(dA), -(dC), -(dG) and -(dT) are used as template with varying efficiencies, with oligo(d)A being the poorest substrate and oligo(dC) the most preferred one. The enzyme is capable of utilising both ribonucleotides and deoxyribonucleotides for primer synthesis in the presence of natural, or synthetic, single-stranded DNA. It has a significantly higher affinity for ribonucleotides than for deoxyribonucleotides. Products with an apparent length of 14 nt and 7 nt are mainly synthesised. In addition to the primase and polymerase activities, the enzyme possesses a template-independent 3'-terminal nucleotidyl transferase activity Saccharolobus solfataricus P2 N(pN)n + n diphosphate
-
 ?

Subunits

Subunits Comment Organism
heterodimer
-
 Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
PriSL
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
 assay at Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 the optimal glycine–NaOH buffer is at pH 9 (pH 8 at 60°C) Saccharolobus solfataricus