DNA polymerase activity of hepatitis B virus particles: differential inhibition by L-enantiomers of nucleotide analogs.
DNA polymerase activity was assayed in hepatitis B virus (HBV) and core particles isolated from chronic producer lines. The particle-associated DNA polymerase activity, which was found to be limited to incorporation of only a few nucleotides, was inhibited by the 5'-triphosphates of nucleoside analogs. The 1-beta-L (1S,4R) and 1-beta-D (1R,4S) enantiomers of antiviral nucleoside analogs were compared for the ability to inhibit incorporation of natural nucleoside triphosphates into the viral DNA. Previously, both enantiomers of several analogs were found to be substrates for human immunodeficiency virus type 1 reverse transcriptase (HIV RT); the 1-beta-D enantiomers of some pairs were preferred as substrates. In contrast, the 1-beta-L enantiomers of all pairs tested were the more potent inhibitors of labeled substrate incorporation into hepatitis B virus DNA; the concentration required to inhibit the incorporation of the natural substrate by 50% was 6-fold to several hundred-fold lower than the concentration of the 1-beta-D enantiomer required for the same inhibitory effect. This preference for the 1-beta-L enantiomers was observed for both RNA-directed synthesis in core particles and DNA-directed synthesis in viral particles. The observed antiviral effect of the nucleoside analogs in cell culture seemed to be limited chiefly by their phosphorylation in cells.
Davis
M G
MG
Glaxo Wellcome Inc., Research Triangle Park, NC 27709, USA.
Wilson
J E
JE
VanDraanen
N A
NA
Miller
W H
WH
Freeman
G A
GA
Daluge
S M
SM
Boyd
F L
FL
Aulabaugh
A E
AE
Painter
G R
GR
Boone
L R
LR
D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003845:Deoxycytosine Nucleotides / Q000378:metabolism; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D000068679:Emtricitabine / Q000031:analogs & derivatives; D006515:Hepatitis B virus / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D007553:Isotope Labeling; D019384:Nucleic Acid Synthesis Inhibitors; D009711:Nucleotides / Q000494:pharmacology*; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000378:metabolism; D016047:Zalcitabine / Q000031:analogs & derivatives / Q000378:metabolism
D004279:DNA, Viral; D003845:Deoxycytosine Nucleotides; D003848:Deoxyguanine Nucleotides; D019384:Nucleic Acid Synthesis Inhibitors; D009711:Nucleotides; D013942:Thymine Nucleotides; C024107:2'-deoxycytidine 5'-triphosphate; D016047:Zalcitabine; C029603:deoxyguanosine triphosphate; D004259:DNA-Directed DNA Polymerase; D000068679:Emtricitabine; C024157:thymidine 5'-triphosphate; C075889:Racivir