title: 2'-Deoxynucleoside 5'-triphosphates modified at alpha-, beta- and gamma-phosphates as substrates for DNA polymerases.
abstract: Replacement of alpha-, beta- and gamma-phosphate groups in 2'-deoxynucleoside 5'-triphosphates (dNTP) with phosphonate groups yields a new set of dNTP mimics with potential biological and therapeutic applications. Here, we describe the synthesis of 15 new dNTPs modified at alpha-, beta- and gamma-phosphates containing, in the case of dUTP, reporter and ligand groups at the C5 position of uracil. It was shown that gamma-substituted dNTPs were substrates for AMV reverse transcriptase despite of the large size of substituent at the gamma-phosphonate. On the other hand, these compounds were poorly utilized by DNA polymerase alpha. For dUTP analogues substituted at both gamma-phosphonate and C5 of uracil, the substrate affinity was 1-2 orders of magnitude lower than for their counterparts containing substituents either at gamma-phosphonate or C5 position. Meanwhile, C5-substituted beta, gamma-dibromomethylenediphosphonates demonstrated poor activity or were not active at all as substrates for AMV reverse transcriptase. Finally, 2'-deoxythymidine 5'-[beta, gamma-(methylphosphinyl)methylphosphonyl]-alpha-phosphate and its 3'-azido-3'-deoxy analog were substrates for AMV reverse transcriptase, but the substrate activity of these analogues was 50-100 times lower as compared with dTTP. HIV reverse transcriptase utilized these compounds 1 order of magnitude less efficiently than AMV reverse transcriptase; terminal deoxynucleotidyl transferase did not recognize them at all.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov str., Moscow 117984, Russia.
authors.forename: L A
authors.initials: LA
authors.lastname: Alexandrova
authors.forename: A Y
authors.initials: AY
authors.lastname: Skoblov
authors.forename: M V
authors.initials: MV
authors.lastname: Jasko
authors.forename: L S
authors.initials: LS
authors.lastname: Victorova
authors.forename: A A
authors.initials: AA
authors.lastname: Krayevsky
mesh_terms: D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D004256:DNA Polymerase I / Q000378:metabolism*; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D006801:Humans; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013379:Substrate Specificity; D013942:Thymine Nucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D014544:Uridine Triphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism
chemical_list: D013942:Thymine Nucleotides; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I; C024157:thymidine 5'-triphosphate; D014544:Uridine Triphosphate