Mechanism-based suppression of dideoxynucleotide resistance by K65R human immunodeficiency virus reverse transcriptase using an alpha-boranophosphate nucleoside analogue

The Journal of Biological Chemistry
B SelmiB Canard

Abstract

The amino acid change K65R in human immunodeficiency virus type 1-reverse transcriptase (RT) confers viral resistance to various 2',3'-dideoxynucleoside drugs in vivo. Using pre-steady state kinetic methods, we found that K65R-reverse transcriptase is 3.2-14-fold resistant to 2',3'-dideoxynucleotides in vitro relative to wild-type reverse transcriptase, in agreement with resistance levels observed in vivo. A decreased catalytic rate constant k(pol) mostly accounts for the lower incorporation efficiency observed for 2',3'-dideoxynucleotides. Examination of the crystal structure of the RT.DNA.dNTP complex suggested that both the charge at position 65 and the 3'-OH of the incoming nucleotide act in synergy during the creation of the phosphodiester bond, resulting in a more pronounced decreased catalytic rate constant for 2',3'-dideoxynucleotides than for dNTPs. This type of intramolecular activation of the leaving phosphate by the 3'-OH group appears to be conserved in several nucleotide phosphotransferases. These data were used to design dideoxynucleotide analogues targeting K65R RT specifically. alpha-Boranophosphate ddATP was found to be a 2-fold better substrate than dATP and inhibited DNA synthesis by K65R RT 153-fold better ...Continue Reading

References

Jan 1, 1984·Cold Spring Harbor Symposia on Quantitative Biology·A M SkalkaJ Leis
Jul 3, 1995·Proceedings of the National Academy of Sciences of the United States of America·S Tabor, C C Richardson
Jan 1, 1995·Methods in Enzymology·F Eckstein, J B Thomson
Aug 16, 1996·The Journal of Biological Chemistry·D ArionM A Parniak
Nov 15, 2000·Cellular and Molecular Life Sciences : CMLS·N Sluis-CremerM A Parniak
Oct 24, 2001·The Journal of Organic Chemistry·Kaizhang HeBarbara Ramsay Shaw

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Citations

Jan 6, 2006·Nucleic Acids Research·Valérie GoldschmidtRoland Marquet
May 1, 2008·PloS One·Scott J GarforthVinayaka R Prasad
Jun 26, 2012·Bioorganic & Medicinal Chemistry Letters·Natsuhisa OkaTakeshi Wada
Apr 9, 2010·Viruses·Kamalendra SinghStefan G Sarafianos
Oct 25, 2014·Viruses·Pinar Iyidogan, Karen S Anderson
Apr 27, 2004·Nature Structural & Molecular Biology·Steve TuskeEddy Arnold
Apr 1, 2011·Future Microbiology·Shiro Ibe, Wataru Sugiura
Apr 2, 2010·Cellular and Molecular Life Sciences : CMLS·Alon Herschhorn, Amnon Hizi
Oct 27, 2005·Nucleosides, Nucleotides & Nucleic Acids·Karine AlvarezBruno Canard
Dec 11, 2007·Nucleosides, Nucleotides & Nucleic Acids·Shoeb I Khan, Barbara Ramsay Shaw
Feb 18, 2010·Protein Science : a Publication of the Protein Society·Pankaj R DagaRobert J Doerksen
Sep 8, 2011·Natural Product Research·S MiladiB Selmi
Oct 27, 2005·Nucleosides, Nucleotides & Nucleic Acids·Mikhail I Dobrikov, Barbara Ramsay Shaw
Jan 31, 2004·Annals of the New York Academy of Sciences·Barbara Ramsay ShawDmitri Sergueev
Oct 27, 2005·Nucleosides, Nucleotides & Nucleic Acids·Shoeb I KhanBarbara Ramsay Shaw
Oct 31, 2015·Future Microbiology·Nicolas Sluis-CremerRaymond F Schinazi
Oct 31, 2007·Chemical Reviews·Ping LiBarbara Ramsay Shaw
Mar 1, 2008·Virus Research·Dirk Jochmans

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