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 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 beta,gamma-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 beta,gamma-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'N3), and ddTMP(3'NH2) from the 3'-end of primers with an equal efficacy.
Kukhanova
Marina K
MK
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov St., Moscow 119991, Russian Federation. kukhan86@hotmail.com
Zakirova
Natalia F
NF
Ivanov
Alexander V
AV
Alexandrova
Ludmila A
LA
Jasco
Maxim V
MV
Khomutov
Alex R
AR
D001483:Base Sequence; D002384:Catalysis; D004247:DNA / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D007700:Kinetics; D015394:Molecular Structure; D010756:Phosphoric Acids / Q000737:chemistry* / Q000378:metabolism*; D010766:Phosphorylation; D013379:Substrate Specificity; D013938:Thymidine Monophosphate / Q000378:metabolism
D017931:DNA Primers; D010756:Phosphoric Acids; D013938:Thymidine Monophosphate; D004247:DNA; D054303:HIV Reverse Transcriptase