Influence of the linkage between leaving group and nucleoside on substrate efficiency for incorporation in DNA catalyzed by reverse transcriptase.
An amino acid deoxyadenosine phosphoramidate and the corresponding phosphodiester congener have been synthesized and tested for primer extension by HIV-1 reverse transcriptase. Replacement of the phosphoramidate linkage of L-histidine-dAMP by a phosphodiester linkage was found to have a beneficial influence on the affinity of this substrate towards HIV-1 reverse transcriptase and on the velocity of the nucleotide incorporation reaction. Modifications of the nature of the P--X bond can be useful to fine-tune the substrate properties of nucleoside triphosphate analogues. Our results also demonstrate that polymerization pausing observed during the incorporation of leaving group modified dNTPs is not caused by the nature of the linkage.
Giraut
Anne
A
Rega Institute Laboratory for Medicinal Chemistry, Catholic University Leuven, Minderborederstraat 10, 3000 Leuven, Belgium.
Herdewijn
Piet
P
D000577:Amides / Q000378:metabolism; D055162:Biocatalysis; D004247:DNA / Q000378:metabolism*; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009705:Nucleosides / Q000378:metabolism*; D010756:Phosphoric Acids / Q000378:metabolism; D013379:Substrate Specificity
D000577:Amides; D003838:Deoxyadenine Nucleotides; D009705:Nucleosides; D010756:Phosphoric Acids; D006639:Histidine; C012873:2'-deoxy-5'-adenosine monophosphate; D004247:DNA; C011067:phosphoramidic acid; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase