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Literature summary for 2.7.7.7 extracted from

  • DeLucia, A.M.; Grindley, N.D.; Joyce, C.M.
    An error-prone family Y DNA polymerase (DinB homolog from Sulfolobus solfataricus) uses a steric gate residue for discrimination against ribonucleotides (2003), Nucleic Acids Res., 31, 4129-4137.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
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Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
F12A mutant ribonucleoside triphosphates almost as efficiently as deoxyribonucleoside triphosphates, and, unlike analogous mutants in other polymerase families, shows no barrier to adding multiple ribonucleotides, suggesting that Dbh can readily accommodate a DNA/RNA duplex product Saccharolobus solfataricus
F12A GTP incorporation by the wild-type enzyme is about 1000fold slower than dGTP incorporation. The rate of GTP incorporation by the mutant enzyme F12A Dbh is 2-3fold slower than incorporation of dGTP. When making a deletion error, ribonucleotide discrimination by wild-type and F12A Dbh is the same as in normal DNA synthesis Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information pre-steady-state kinetic constants for dNTP and rNTP incorporation by wild-type and F12A mutant enzyme Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
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-
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Saccharolobus solfataricus P96022
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-

Purification (Commentary)

Purification (Comment) Organism
wild-type and F12A mutant Saccharolobus solfataricus
wild-type and mutant enzyme F12A Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deoxynucleoside triphosphate + DNAn ribonucleotide discrimination by the DinB homolog (Dbh) DNA polymerase is as stringent as in other polymerases. When making a deletion error, ribonucleotide discrimination by wild-type and F12A Dbh is the same as in normal DNA synthesis Saccharolobus solfataricus diphosphate + DNAn+1
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?
deoxynucleoside triphosphate + DNAn GTP incorporation by the wild-type enzyme is about 1000fold slower than dGTP incorporation. The rate of GTP incorporation by the mutant enzyme F12A Dbh is 2-3fold slower than incorporation of dGTP. The enzyme makes single-base deletion errors at high frequency in particular sequence contexts Saccharolobus solfataricus diphosphate + DNAn+1
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?

Synonyms

Synonyms Comment Organism
Dbh DNA polymerase
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Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information pre-steady-state kinetic constants for dNTP and rNTP incorporation by wild-type and F12A mutant enzyme Saccharolobus solfataricus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
additional information pre-steady-state kinetic constants for dNTP and rNTP incorporation by wild-type and F12A mutant enzyme Saccharolobus solfataricus