S-Triphosphates of 1-(2',3'-epithio-2'.3'-dideoxy-B_D-lyxofuranosyl)thymine, l-(2',3'-eptthio-~,~-dideoxy-~-D-ribofuranosyl)th~lne and 2',3'-lyxoanhydrothymidine have been shown to be termmatton substrates for human immunodeficiency virus (HIV) and avian myeloblastosis virus (AMV) reverse transcriptases as well as DNA polymerase 1 from E. co11 and DNA polymerase j3 from rat hver. At the same time they do not terminate DNA synthesis catalysed by DNA polymerase E from human placenta. Km values of ItTTP. rtTTP and laTTP incorporation mto the DNA chain during catalysts by AMV reverse transcriptase agree closely with each other being 1.5-2.5 times higher than K,,, value for dTTP Furthermore, V,,, values for modified substrates are only 2-3 times lower than b',,,',,, for dTTP. The evidence favours the hypothesis of high affinity of modified nucleotides wtth a flattened furanosyl rmg for DNA polymerase active sites.DNA polymerase: Terminatton substrate; Modified nucleoside S-trtphosphate