5'-derivatives of oligonucleotides as primers of DNA polymerization catalyzed by AMV reverse transcriptase and Klenow fragment of DNA polymerase 1

FEBS Lett. 1991 Apr 9;281(1-2):111-3. doi: 10.1016/0014-5793(91)80371-9.

Abstract

The Km and Vmax values for d(pT)8 and its derivatives containing various 5'-end groups were estimated in the reaction of polymerization catalyzed with AMV-RT and FK. The change in affinity of modified primers was more pronounced in the case of AMV-RT than in the case of FK. Introducing in d(pT)8 of intercalators such as phenazinium, ethidium and daunomycin residues results in 2.7-, 8.7- and 11-fold increases in the primer affinity to AMV-RT, respectively. However, in the case of hemin and cholesterol derivatives the Km values were 3 and 5 times higher than those for d(pT)8. Compared to d(pT)8, the affinity of FK to all the above analogs was 2.3-3.6 times higher with the exception of cholesterol derivative to which it was 2.4-fold lower. The effect of the 5'-end residues on the Vmax values of d(pT)8 was small and ranged from 44% to 120% of that for d(pT)8. Therefore such reactive derivatives of oligonucleotides can be used as effective primers of AMV-RT and FK. Possible reasons for various effects of the 5'-end residues of the primer on its interaction with FK or AMV-RT in the presence of poly(A) are discussed.

MeSH terms

  • Avian Myeloblastosis Virus / enzymology*
  • DNA Polymerase I / metabolism*
  • DNA Replication*
  • Kinetics
  • Oligonucleotide Probes / chemical synthesis
  • Oligonucleotide Probes / metabolism*
  • Peptide Fragments / metabolism
  • RNA-Directed DNA Polymerase / metabolism*
  • Structure-Activity Relationship

Substances

  • Oligonucleotide Probes
  • Peptide Fragments
  • RNA-Directed DNA Polymerase
  • DNA Polymerase I