title: A conserved African swine fever virus IkappaB homolog, 5EL, is nonessential for growth in vitro and virulence in domestic swine.
abstract: An African swine fever virus (ASFV) gene with similarity to the cellular inhibitor of NFkappaB (IkappaB) was described in the pathogenic African isolate Malawi Lil-20/1 (ORF 5EL) and a cell-culture-adapted European virus, BA71V (ORF A238L). Recently, this gene was shown to be a functional IkappaB homolog capable of downregulating NFkappaB-regulated gene expression. This observation suggests the gene may be of significance to aspects of ASFV pathogenesis and virulence in domestic swine by interfering with a normal antiviral host response. Here we show, using nucleotide sequence analysis, that 5EL is highly conserved among various African and European pathogenic field isolates and that in all cases its similarity to IkappaB genes is limited to the presence of four low complexity ankyrin repeats in the ASFV gene. The 5EL gene of Malawi Lil-20/1 encodes a 28-kDa protein which was expressed early in virus-infected macrophage cell cultures with maximum levels observed at 3 to 5 hr postinfection. To study gene function, a Malawi Lil-20/1 5EL gene deletion mutant (Delta5EL) was constructed. Growth characteristics of Delta5EL in porcine macrophage cell cultures were indistinguishable from those of the parental virus. And, Delta5EL exhibited an unaltered parental Malawi Lil-20/1 disease and virulence phenotype in domestic swine. Thus, although highly conserved among ASFV isolates, 5EL is nonessential for growth in porcine macrophages in vitro and for viral virulence in domestic swine. A possible role for this gene in transmission of ASFV in nature, a setting which involves the cycling of ASFV between two highly adapted hosts, Ornithodoros ticks and warthogs or bush pigs, in sub-Saharan Africa is discussed.
authors.affiliation: Agricultural Research Service, U.S. Department of Agriculture, Greenport, New York, 11944-0848, USA.
authors.forename: J G
authors.initials: JG
authors.lastname: Neilan
authors.forename: Z
authors.initials: Z
authors.lastname: Lu
authors.forename: G F
authors.initials: GF
authors.lastname: Kutish
authors.forename: L
authors.initials: L
authors.lastname: Zsak
authors.forename: T L
authors.initials: TL
authors.lastname: Lewis
authors.forename: D L
authors.initials: DL
authors.lastname: Rock
mesh_terms: D000357:African Swine Fever / Q000401:mortality; D000358:African Swine Fever Virus / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity*; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D018562:Disease Transmission, Infectious / Q000662:veterinary; D008264:Macrophages / Q000821:virology; D008969:Molecular Sequence Data; D009154:Mutation; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D015996:Survival Rate; D013552:Swine; D013997:Time Factors; D051841:Transcription Factor RelB; D014157:Transcription Factors; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: C108479:5EL protein, African swine fever virus; D011518:Proto-Oncogene Proteins; D014157:Transcription Factors; D014764:Viral Proteins; D051841:Transcription Factor RelB