title: Functional characterization and inhibition of the type II DNA topoisomerase coded by African swine fever virus.
abstract: DNA topoisomerases are essential for DNA metabolism and while their role is well studied in prokaryotes and eukaryotes, it is less known for virally-encoded topoisomerases. African swine fever virus (ASFV) is a nucleo-cytoplasmic large DNA virus that infects Ornithodoros ticks and all members of the family Suidae, representing a global threat for pig husbandry with no effective vaccine nor treatment. It was recently demonstrated that ASFV codes for a type II topoisomerase, highlighting a possible target for control of the virus. In this work, the ASFV DNA topoisomerase II was expressed in Saccharomyces cerevisiae and found to efficiently decatenate kDNA and to processively relax supercoiled DNA. Optimal conditions for its activity were determined and its sensitivity to a panel of topoisomerase poisons and inhibitors was evaluated. Overall, our results provide new knowledge on viral topoisomerases and on ASFV, as well as a possible target for the control of this virus.
authors.affiliation: CIISA, Faculdade de Medicina Veterinária, ULisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal. Electronic address: jnscoelho@gmail.com.
authors.forename: João
authors.initials: J
authors.lastname: Coelho
authors.affiliation: CIISA, Faculdade de Medicina Veterinária, ULisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal.
authors.forename: Fernando
authors.initials: F
authors.lastname: Ferreira
authors.affiliation: CIISA, Faculdade de Medicina Veterinária, ULisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal.
authors.forename: Carlos
authors.initials: C
authors.lastname: Martins
authors.affiliation: CIISA, Faculdade de Medicina Veterinária, ULisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal. Electronic address: alexandre@fmv.ulisboa.pt.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Leitão
mesh_terms: D000358:African Swine Fever Virus / Q000201:enzymology* / Q000235:genetics; D049933:Aminocoumarins / Q000494:pharmacology; D000677:Amsacrine / Q000494:pharmacology; D016831:Crithidia fasciculata / Q000235:genetics; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D004317:Doxorubicin / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics; D059005:Topoisomerase II Inhibitors / Q000494:pharmacology*
chemical_list: D049933:Aminocoumarins; D059005:Topoisomerase II Inhibitors; D000677:Amsacrine; D004317:Doxorubicin; D004250:DNA Topoisomerases, Type II; C004628:coumermycin
keywords: ASFV; African swine fever virus; Amsacrine; Ciprofloxacin; DNA Topoisomerases, Type II; Doxorubicin; P1192R; Topoisomerase II inhibitors