Hypophosphoric acid is a unique substrate of pyrophosphorolysis catalyzed by HIV-1 reverse transcriptase

Pyrophosphate analogues, namely, pyrophosphorous, hypophosphoric, and hypophosphorous acids, were evaluated as inhibitors in elongation reactions and substrates in pyrophosphorolysis reaction catalyzed by HIV-1 reverse transcriptase and DNA polymerase I (the Klenow fragment). The substrate efficacy...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2005-12, Vol.338 (3), p.1335-1341
Main Authors: Kukhanova, Marina K., Zakirova, Natalia F., Ivanov, Alexander V., Alexandrova, Ludmila A., Jasco, Maxim V., Khomutov, Alex R.
Format: Article
Language:English
Subjects:
Base Sequence
Catalysis
DNA - genetics
DNA - metabolism
DNA Primers - genetics
HIV Reverse Transcriptase - metabolism
HIV-1 reverse transcriptase
Human immunodeficiency virus 1
Kinetics
Klenow fragment
Molecular Structure
Phosphoric Acids - chemistry
Phosphoric Acids - metabolism
Phosphorylation
Pyrophosphate analogues
Pyrophosphorolysis
Substrate Specificity
Thymidine Monophosphate - metabolism
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Summary:Pyrophosphate analogues, namely, pyrophosphorous, hypophosphoric, and hypophosphorous acids, were evaluated as inhibitors in elongation reactions and substrates in pyrophosphorolysis reaction catalyzed by HIV-1 reverse transcriptase and DNA polymerase I (the Klenow fragment). The substrate efficacy of hypophosphoric acid in pyrophosphorolysis reaction exceeded that of pyrophosphate for both enzymes by more than ten times. The product of the reaction was a dNTP analogue bearing a hypophosphate in the β,γ-position. Pyrophosphorous and hypophosphorous acids were neither inhibitors nor substrates for the enzymes. Kinetic parameters of the pyrophosphorolysis reaction catalyzed by HIV reverse transcriptase in the presence of hypophosphoric acid were evaluated. The dTMP analogue bearing a hypophosphate in the β,γ-position was synthesized and its substrate properties in elongation reaction catalyzed by HIV-1 reverse transcriptase were similar to those of natural dTTP. Hypophosphoric acid was capable of removing ddTMP, ddTMP(3′N 3), and ddTMP(3′NH 2) from the 3′-end of primers with an equal efficacy.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2005.10.092