Incorporation of 4-thiothymidine into DNA by the Klenow fragment and HIV-1 reverse transcriptase

Bioorg Med Chem Lett. 2000 May 1;10(9):907-10. doi: 10.1016/s0960-894x(00)00123-2.

Abstract

The 5'-triphosphate of 4-thiothymidine (4S-TTP) is an excellent substrate for the Klenow fragment of Escherichia coli DNA polymerase 1 and HIV-1 reverse transcriptase with values of k(cat)/Km within a factor of approximately 3 of those for TTP. A large UV change (deltaepsilon= -9770 M(-1)cm(-1) at 340 nm) associated with incorporation of 4S-TMP into nucleic acid duplexes makes possible a rapid, continuous spectrophotometric assay of the reaction progress.

MeSH terms

  • Base Pairing / drug effects
  • DNA / biosynthesis*
  • DNA / chemistry
  • DNA Polymerase I / metabolism*
  • DNA Primers
  • Escherichia coli / enzymology
  • HIV Reverse Transcriptase / metabolism*
  • Kinetics
  • Spectrophotometry, Ultraviolet
  • Templates, Genetic
  • Thymidine / analogs & derivatives*
  • Thymidine / chemistry

Substances

  • 4-thiothymidine
  • DNA Primers
  • DNA
  • HIV Reverse Transcriptase
  • DNA Polymerase I
  • Thymidine