In vitro mutagenesis of the herpes simplex virus type 1 DNA polymerase gene results in altered drug sensitivity of the enzyme.
A mutation (asparagine 815 to serine 815) was introduced into the herpes simplex virus type 1 (HSV-1) DNA polymerase (pol). The HSV-1 pol enzyme in lysates of Saccharomyces cerevisiae cells expressing the mutant protein showed increased resistance to acyclovir triphosphate and increased sensitivity to phosphonoacetate but was not substantially altered with respect to sensitivity to phosphonoformate or aphidicolin. These results directly demonstrate that both resistance to acyclovir triphosphate and sensitivity to phosphonoacetate can be conferred by this mutation in the absence of other viral factors and that the yeast expression system can be used for structure-function studies on HSV-1 pol.
Matthews
J T
JT
Department of Virology, Squibb Institute for Medical Research, Princeton, New Jersey 08543-4000.
Carroll
R D
RD
Stevens
J T
JT
Haffey
M L
ML
D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D016590:Aphidicolin; D001216:Asparagine; D001483:Base Sequence; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D004224:Diterpenes / Q000494:pharmacology*; D017245:Foscarnet; D005814:Genes, Viral; D005822:Genetic Vectors; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D019384:Nucleic Acid Synthesis Inhibitors; D015345:Oligonucleotide Probes; D010746:Phosphonoacetic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012694:Serine; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics*
D000998:Antiviral Agents; D004224:Diterpenes; D019384:Nucleic Acid Synthesis Inhibitors; D015345:Oligonucleotide Probes; D015678:Viral Structural Proteins; D017245:Foscarnet; D016590:Aphidicolin; D012694:Serine; D001216:Asparagine; D004259:DNA-Directed DNA Polymerase; D010746:Phosphonoacetic Acid