African swine fever virus AP endonuclease is a redox-sensitive enzyme that repairs alkylating and oxidative damage to DNA.
African swine fever virus (ASFV) encodes an AP endonuclease (pE296R) which is essential for virus growth in swine macrophages. We show here that the DNA repair functions of pE296R (AP endonucleolytic, 3'-->5' exonuclease, 3'-diesterase and nucleotide incision repair (NIR) activities) and DNA binding are inhibited by reducing agents. Protein pE296R contains one intramolecular disulfide bond, whose disruption by reducing agents might perturb the interaction of the viral AP endonuclease with the DNA substrate. The characterization of the 3'-->5' exonuclease and 3'-repair diesterase activities of pE296R indicates that it has strong preference for mispaired and oxidative base lesions at the 3'-termini of single-strand breaks. Finally, the viral protein protects against DNA damaging agents in both prokaryotic and eukaryotic cells, emphasizing its importance in vivo. The biochemical and genetic properties of ASFV AP endonuclease are consistent with the repair of DNA damage generated by the genotoxic intracellular environment of the host macrophage.
Redrejo-Rodríguez
Modesto
M
Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas/Universidad Autónoma de Madrid), Universidad Autónoma de Madrid, C/Nicolás Cabrera 1, 28049 Madrid, Spain.
Ishchenko
Alexander A
AA
Saparbaev
Murat K
MK
Salas
María L
ML
Salas
José
J
D000358:African Swine Fever Virus / Q000201:enzymology* / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology*; D004279:DNA, Viral / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005814:Genes, Viral; D005816:Genetic Complementation Test; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000502:physiology; D066298:In Vitro Techniques; D008264:Macrophages / Q000378:metabolism / Q000821:virology; D009153:Mutagens / Q000494:pharmacology; D010084:Oxidation-Reduction; D019163:Reducing Agents / Q000494:pharmacology; D014709:Vero Cells
D004279:DNA, Viral; D009153:Mutagens; D019163:Reducing Agents; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D004229:Dithiothreitol