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HIV-1 reverse transcriptase: inhibition by 2',5'-oligoadenylates.

R W Sobol·W L Fisher·N L Reichenbach·A Kumar·W A Beard·S H Wilson·R Charubala·W Pfleiderer·R J Suhadolnik
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Abstract

2',5'-Oligoadenylates (2-5A) and derivatives are noncompetitive inhibitors of primer/HIV-1 reverse transcriptase complex formation. The mechanism and specificity of this inhibitory action of 2-5A and 2-5A derivatives have been evaluated with 2-5A molecules modified in ribosyl moiety, chain length, extent of 5'-phosphorylation, and 2',5'-phosphodiester linkage. UV covalent cross-linking of preformed complexes of p66/p66 homodimer or p66/p51 heterodimer recombinant HIV-1 reverse transcriptase and the primer analog pd(T)16 allowed analysis of the initial step in HIV-1 reverse transcriptase-catalyzed DNA synthesis. Utilizing this primer binding assay, it is demonstrated that 2-5A and 2-5A derivatives inhibit the binding of pd(T)16 to HIV-1 reverse transcriptase. This inhibition is specific for the 2',5'-internucleotide linkage in that the corresponding 3',5'-adenylate derivatives do not exhibit inhibitory activity. Enhanced inhibitory properties were observed following modifications of the 2-5A molecule which result in an increase in hydrophobicity. Replacement of the D-ribosyl moiety of 2-5A with the 3'-deoxyribosyl moiety increased the inhibition of primer/HIV-1 reverse transcriptase complex formation 15-20%. 2',5'-Phosphorothioate substitution yielded the most effective inhibitors, with Ki's of 7-13 microM. In all cases, inhibition of primer/HIV-1 reverse transcriptase complex formation showed a preference for the 5'-triphosphate moiety. Nonphosphorylated derivatives were not inhibitory; 5'-monophosphate derivatives exhibited little or no inhibition. The inhibition of primer binding to HIV-1 reverse transcriptase correlated well with the inhibition of DNA-directed DNA synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

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Adenine NucleotidesAntiviral AgentsDNAHIV Reverse TranscriptaseHIV-1OligoribonucleotidesPhosphorylationPhotochemistryReverse Transcriptase InhibitorsThymidineJournal ArticleResearch Support, U.S. Gov't, Non-P.H.S.

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R W Sobol, W L Fisher, N L Reichenbach, A Kumar, W A Beard, S H Wilson, R Charubala, W Pfleiderer, R J Suhadolnik. 1993-11-16. HIV-1 reverse transcriptase: inhibition by 2',5'-oligoadenylates.. https://doi.org/10.1021/bi00096a023

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