African swine fever virus-induced DNA polymerase is resistant to aphidicolin.
African swine fever virus (ASFV) induces the synthesis of a virus-specific DNA polymerase, which is inhibited by phosphonoacetic acid and cytosine arabinoside. In contrast to all other alpha-like DNA polymerases of DNA viruses, ASFV-specific DNA polymerase is resistant to aphidicolin. Concentrations of the drug as high as 160 microM had no effect on virus production or plaquing efficiency. The resistance of ASFV DNA polymerase to aphidicolin was confirmed by analyzing the effect of the drug on viral DNA synthesis. A moderate inhibition of viral DNA synthesis was observed when aphidicolin was added immediately after virus adsorption but normal synthesis occurred, with a peak at 10 hr p.i., when the drug was added at 2 or 4 hr p.i. This suggests that a very early phase of ASFV DNA replication is sensitive to aphidicolin and is probably catalyzed by a different enzyme. An in vitro assay of DNA polymerase activity was used to assay the sensitivity of the virus-specific DNA polymerase to inhibitors. In correspondence to the results observed in vivo, phosphonoacetic acid strongly inhibited the enzyme activity, whereas aphidicolin had no effect. Resistance to aphidicolin was independent of the concentration of dCTP used in the assay. Three independent ASFV mutants resistant to phosphonoacetic acid showed the same resistance to aphidicolin as wild type virus.
Marques
M I
MI
Gulbenkian Institute of Science, Oeiras, Portugal.
Costa
J V
JV
D000358:African Swine Fever Virus / Q000187:drug effects / Q000201:enzymology*; D016590:Aphidicolin / Q000494:pharmacology*; D003561:Cytarabine / Q000494:pharmacology; D004279:DNA, Viral / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000187:drug effects* / Q000378:metabolism; D004352:Drug Resistance, Microbial; D007700:Kinetics; D010746:Phosphonoacetic Acid / Q000494:pharmacology
D004279:DNA, Viral; D003561:Cytarabine; D016590:Aphidicolin; D004259:DNA-Directed DNA Polymerase; D010746:Phosphonoacetic Acid