Comparison of initiating abilities of primers of different length in polymerization reactions catalyzed by DNA polymerases from thermoacidophilic archaebacteria

Biochim Biophys Acta. 1989 Jun 1;1008(1):102-7. doi: 10.1016/0167-4781(89)90175-9.


Optimal conditions for polymerization reaction catalyzed on poly(dA) and poly(dT) templates by DNA polymerases from thermoacidophilic archaebacteria--DNA polymerase A from Sulfolobus acidocaldarius and DNA polymerase B from Thermoplasma acidophilum--have been established. Values of Km and Vmax (60 degrees C) for a set of primers d(pA)n and d(pT)n have been estimated. Minimal primers for both enzymes are dNMP. Lengthening of primers by each mononucleotide increases their affinity about 2.16-fold. Linear dependence of log Km and of log vmax on the number of mononucleotide links in primers (n) has breaking point at n = 10. The value of Vmax is about 20% of that for decanucleotide. The affinity of the primer d(pA)9p(rib*) with a deoxyribosylurea residue at the 3'-end does not differ essentially from that of d(pA)9. Substitution of the 3'-terminal nucleotide of a complementary primer for a noncomplementary nucleotide, e.g., substitution of 3'-terminal A for C in d(pA)10 in the reaction catalyzed on poly(dT), decreases the affinity of a primer by one order of magnitude.

Publication types

  • Comparative Study

MeSH terms

  • Archaea / enzymology*
  • Bacteria / enzymology*
  • DNA-Directed DNA Polymerase / metabolism*
  • Kinetics
  • Poly A
  • Poly T
  • Poly dA-dT
  • Structure-Activity Relationship
  • Templates, Genetic


  • Poly A
  • Poly T
  • poly(dA)
  • Poly dA-dT
  • DNA-Directed DNA Polymerase