Functional and molecular characterization of African swine fever virus mutants resistant to phosphonoacetic acid.

Abstract:

:African swine fever virus (ASFV) growth and plaque formation were inhibited by phosphonoacetic acid (PAA) concentrations of 200 micrograms/ml or more. One spontaneous mutant and two mutants isolated from mutagenized virus were resistant to PAA inhibition and showed practically normal viral DNA synthesis in the presence of PAA. DNA polymerase activity present in the cytoplasmic fraction from cells infected with the mutants required 10-fold higher concentrations of PAA for inhibition compared to equivalent inhibition of the wild-type enzyme. Like wild-type virus, the PAA-resistant mutants were resistant to inhibition by aphidicolin. Marker rescue analysis with mutant DNA fragments covering different regions of the ASFV DNA polymerase gene mapped the mutations within a fragment which was cloned and sequenced. A single nucleotide and amino acid change was assigned to each mutant. Two of the PAA-resistant mutations lie within the highly conserved region II common to alpha-like DNA polymerases, which has been implicated in pyrophosphate binding and probably also in dNTP binding. The other mutation was localized to within a region of moderate homology among viral DNA polymerases close to one of the motifs allegedly considered as constituting the 3'-5' exonuclease active site.

journal_name

Virology

journal_title

Virology

authors

Marques MI,Costa JV

doi

10.1006/viro.1995.9953

subject

Has Abstract

pub_date

1995-12-01 00:00:00

pages

72-81

issue

1

eissn

0042-6822

issn

1096-0341

pii

S0042682285799539

journal_volume

214

pub_type

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