Chiral discrimination of enantiomeric 2'-deoxythymidine 5'-triphosphate by HIV-1 reverse transcriptase and eukaryotic DNA polymerases.
Inhibitory effects of 2'-deoxy-L-thymidine 5'-triphosphate (L-dTTP), the enantiomer of the natural substrate D-dTTP, on the activity of mammalian DNA polymerases alpha, beta and gamma, Escherichia coli DNA polymerase I and human immunodeficiency virus 1 (HIV-1) reverse transcriptase were examined. When poly(rA)n-oligo(dT)12-18 was used as the template-primer, L-dTTP showed remarkable inhibitory effect on HIV-1 reverse transcriptase in competitive fashion with respect to the substrate dTTP. In contrast, L-dTTP did not inhibit DNA polymerases alpha and was slightly inhibitory to DNA polymerase beta. These results suggest that the nuclear DNA polymerases alpha and beta showed high specificity for the substrate with the natural configuration of the sugar moiety, D-dTTP, exhibiting little or no ability to recognize L-dTTP, whereas HIV-1 reverse transcriptase essentially lacked the ability to differentiate the D- and L-sugar moieties.
Yamaguchi
T
T
Department of Biological Sciences, Nishi-Tokyo University, Yamanashi, Japan.
Iwanami
N
N
Shudo
K
K
Saneyoshi
M
M
D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D004256:DNA Polymerase I / Q000037:antagonists & inhibitors / Q000378:metabolism; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology*; D066298:In Vitro Techniques; D007700:Kinetics; D019384:Nucleic Acid Synthesis Inhibitors; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D051381:Rats; D018894:Reverse Transcriptase Inhibitors; D013237:Stereoisomerism; D013379:Substrate Specificity; D013942:Thymine Nucleotides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
D019384:Nucleic Acid Synthesis Inhibitors; D009838:Oligodeoxyribonucleotides; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D011061:Poly A; C018368:poly(rA).oligo(dT); D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase; C024157:thymidine 5'-triphosphate