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

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

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)...

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Veröffentlicht in:Bioorganic & medicinal chemistry letters 2000-05, Vol.10 (9), p.907-910
Hauptverfasser: Rao, T V, Haber, M T, Sayer, J M, Jerina, D M
Format: Artikel
Sprache:eng
Schlagworte:
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
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Zusammenfassung: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.
ISSN:0960-894X
DOI:10.1016/S0960-894X(00)00123-2