title: African swine fever virus ORF P1192R codes for a functional type II DNA topoisomerase.
abstract: Topoisomerases modulate the topological state of DNA during processes, such as replication and transcription, that cause overwinding and/or underwinding of the DNA. African swine fever virus (ASFV) is a nucleo-cytoplasmic double-stranded DNA virus shown to contain an OFR (P1192R) with homology to type II topoisomerases. Here we observed that pP1192R is highly conserved among ASFV isolates but dissimilar from other viral, prokaryotic or eukaryotic type II topoisomerases. In both ASFV/Ba71V-infected Vero cells and ASFV/L60-infected pig macrophages we detected pP1192R at intermediate and late phases of infection, cytoplasmically localized and accumulating in the viral factories. Finally, we used a Saccharomyces cerevisiae temperature-sensitive strain in order to demonstrate, through complementation and in vitro decatenation assays, the functionality of P1192R, which we further confirmed by mutating its predicted catalytic residue. Overall, this work strengthens the idea that P1192R constitutes a target for studying, and possibly controlling, ASFV transcription and replication.
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. Electronic address: cmartins@fmv.ulisboa.pt.
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: fernandof@fmv.ulisboa.pt.
authors.forename: Fernando
authors.initials: F
authors.lastname: Ferreira
authors.affiliation: Instituto de Investigação Científica Tropical, CVZ, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal; CIISA, 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: D000357:African Swine Fever / Q000821:virology; D000358:African Swine Fever Virus / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D004250:DNA Topoisomerases, Type II / Q000235:genetics* / Q000378:metabolism*; D005816:Genetic Complementation Test; D008264:Macrophages / Q000821:virology; D016366:Open Reading Frames; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D034421:Sus scrofa; D013552:Swine; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014764:Viral Proteins; D004250:DNA Topoisomerases, Type II
keywords: African swine fever virus (ASFV); P1192R; Type II DNA topoisomerase