Inhibition of reverse transcriptase from feline immunodeficiency virus by analogs of 2′-deoxyadenosine-5′-triphosphate

The replication of feline immunodeficiency virus (FIV) in cultured cells was inhibited by 2′,3′-dideoxyadenosine (ddA) and by 9-(2-phosphonylmethoxyethyI)adenine (PMEA) with IC₅₀ values of 0.98 and 0.95 μM, respectively. The effects of the presumed active forms of these inhibitors, ddATP and PMEA-diphosphate (PMEApp), upon the FIV reverse transcriptase (RT) were examined with two different template-primer systems. Both of these compounds were potent inhibitors of the FIV RT in reactions with primed φX-174 DNA, yielding K_i values of 8.8 nM for ddATP and 5.0 nM for PMEApp. However, they were both poor inhibitors of the reaction with poly(rU)-oligo(dA); concentrations of ddATP or PMEApp greater than 10 μM were required to inhibit this reaction by 50%. Further analysis of the reaction with poly(rU)-oligo(dA) revealed that even in the absence of inhibitors the primers were extended by less than 20 nucleotides. In contrast, high molecular weight products were obtained in reactions with φX-174 DNA. These results suggest that the reaction of FIV RT with poly(rU)-oligo(dA) is not highly processive. The high degree of termination encountered during this reaction with poly(rU)-oligo(dA) may be responsible for the low inhibitory potential of ddATP and PMEApp.