[Formation of phosphonoester bonds, catalyzed by DNA polymerases]

Mol Biol (Mosk). 1991 Nov-Dec;25(6):1688-700.
[Article in Russian]

Abstract

3'-Fluoro-2',3'-dideoxythymidine 5'-(alpha-methylphosphonyl)-beta, gamma-diphosphate (I) and 2'-deoxythymidine 5'-(alpha-methylphosphonyl)-beta,gamma-diphosphate (II) were synthesised. Reverse transcriptases of HIV and avian myeloblastosis virus, rat liver DNA polymerase beta, calf thymus terminal deoxynucleotidyl transferase and E. coli DNA polymerase I KF incorporated both compounds into the growing DNA chain, KF being the least effective. Compound I revealed termination substrate properties, but II was repeatedly incorporated into the DNA chain, for example, by HIV reverse transcriptase - up to 8 residues. Human placenta DNA polymerases alpha and epsilon incorporated neither I nor II into the DNA chain, although DNA synthesis, catalyzed by all the investigated enzymes, was inhibited in the presence of I or II and compound II was a more effective inhibitor then I. The DNA fragments containing alpha-phosphonomethyl groups were hydrolyzed by 3'----5' exonuclease of DNA polymerase I and not hydrolyzed by ExoIII from E. coli.

Publication types

  • English Abstract

MeSH terms

  • Animals
  • Autoradiography
  • Avian Myeloblastosis Virus / enzymology
  • Base Sequence
  • Catalysis
  • Cattle
  • DNA-Directed DNA Polymerase / metabolism*
  • Dideoxynucleosides / antagonists & inhibitors
  • Dideoxynucleosides / chemical synthesis
  • Dideoxynucleosides / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • HIV / enzymology
  • Humans
  • Liver / enzymology
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Placenta / enzymology
  • Pregnancy
  • RNA-Directed DNA Polymerase / metabolism
  • Rats
  • Thymine Nucleotides / antagonists & inhibitors
  • Thymine Nucleotides / chemical synthesis
  • Thymine Nucleotides / chemistry
  • Thymus Gland / enzymology

Substances

  • Dideoxynucleosides
  • Thymine Nucleotides
  • 2'-deoxythymidine-5'-(alpha-methylphosphonyl)-beta,gamma-diphosphate
  • 3'-fluoro-2',3'-dideoxythymidine-5'-(alpha-methylphosphonyl)-beta,gamma-diphosphate
  • RNA-Directed DNA Polymerase
  • DNA-Directed DNA Polymerase