2',3'-Dideoxy-3'-thionucleoside triphosphates: syntheses and polymerase substrate activities

Org Lett. 2007 Mar 15;9(6):1161-3. doi: 10.1021/ol070147w. Epub 2007 Feb 24.

Abstract

All four 2',3'-dideoxy-3'-thio-nucleosides (ddtNTPs) function as substrates for the Y410F mutant of Deep Vent (exo-) DNA polymerase. Not only are the ddtNTPs incorporated to form the N + 1 product, but further elongations are observed in which the key step is attack of the 3'-thiol on the 5'-triphosphate. Although other polymerases are likely to differ in their use of the ddtNTPs, there does not appear to be a fundamental prohibition against using a thiol nucleophile on a phosphate anhydride electrophile. The syntheses of four ddtNTPs (C, T, A, G) are described. [structure: see text]

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Catalysis
  • DNA-Directed DNA Polymerase / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors / chemical synthesis*
  • Enzyme Inhibitors / pharmacology*
  • Mutation
  • Nucleic Acid Synthesis Inhibitors*
  • Phosphates / chemistry
  • Substrate Specificity
  • Sulfhydryl Compounds / chemistry
  • Thionucleosides / chemical synthesis*

Substances

  • Enzyme Inhibitors
  • Nucleic Acid Synthesis Inhibitors
  • Phosphates
  • Sulfhydryl Compounds
  • Thionucleosides
  • Deep Vent DNA polymerase
  • DNA-Directed DNA Polymerase