The availability of highly purified recombinant enzymes and model heteropolymeric nucleic acid substrates now allows more precise evaluation of the ribonuclease H (RNase H) activity associated with human immunodeficiency virus (HIV) reverse transcriptase. In addition to degrading the RNA–DNA replicative intermediate, this C-terminal domain of around 130 residues supports highly specialized events that cannot be complemented by host-coded enzymes during retrovirus replication. RNase H activity should therefore be considered a plausible candidate for therapeutic intervention. Events during HIV replication requiring precise RNase H-mediated hydrolysis, the methodologies available to study these events, and their potential for therapeutic intervention are reviewed here.
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