title: Biological characterization and next-generation genome sequencing of the unclassified Cotia virus SPAn232 (Poxviridae).
abstract: Cotia virus (COTV) SPAn232 was isolated in 1961 from sentinel mice at Cotia field station, São Paulo, Brazil. Attempts to classify COTV within a recognized genus of the Poxviridae have generated contradictory findings. Studies by different researchers suggested some similarity to myxoma virus and swinepox virus, whereas another investigation characterized COTV SPAn232 as a vaccinia virus strain. Because of the lack of consensus, we have conducted an independent biological and molecular characterization of COTV. Virus growth curves reached maximum yields at approximately 24 to 48 h and were accompanied by virus DNA replication and a characteristic early/late pattern of viral protein synthesis. Interestingly, COTV did not induce detectable cytopathic effects in BSC-40 cells until 4 days postinfection and generated viral plaques only after 8 days. We determined the complete genomic sequence of COTV by using a combination of the next-generation DNA sequencing technologies 454 and Illumina. A unique contiguous sequence of 185,139 bp containing 185 genes, including the 90 genes conserved in all chordopoxviruses, was obtained. COTV has an interesting panel of open reading frames (ORFs) related to the evasion of host defense, including two novel genes encoding C-C chemokine-like proteins, each present in duplicate copies. Phylogenetic analysis revealed the highest amino acid identity scores with Cervidpoxvirus, Capripoxvirus, Suipoxvirus, Leporipoxvirus, and Yatapoxvirus. However, COTV grouped as an independent branch within this clade, which clearly excluded its classification as an Orthopoxvirus. Therefore, our data suggest that COTV could represent a new poxvirus genus.
authors.affiliation: Laboratório de Biologia Molecular de Vírus, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Priscila P
authors.initials: PP
authors.lastname: Afonso
authors.forename: Patrícia M
authors.initials: PM
authors.lastname: Silva
authors.forename: Laila C
authors.initials: LC
authors.lastname: Schnellrath
authors.forename: Desyreé M
authors.initials: DM
authors.lastname: Jesus
authors.forename: Jianhong
authors.initials: J
authors.lastname: Hu
authors.forename: Yajie
authors.initials: Y
authors.lastname: Yang
authors.forename: Rolf
authors.initials: R
authors.lastname: Renne
authors.forename: Marcia
authors.initials: M
authors.lastname: Attias
authors.forename: Richard C
authors.initials: RC
authors.lastname: Condit
authors.forename: Nissin
authors.initials: N
authors.lastname: Moussatché
authors.forename: Clarissa R
authors.initials: CR
authors.lastname: Damaso
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D003429:Cross Reactions / Q000276:immunology; D003588:Cytopathogenic Effect, Viral; D005814:Genes, Viral; D016679:Genome, Viral; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008253:Macaca mulatta; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010802:Phylogeny; D011212:Poxviridae / Q000145:classification* / Q000235:genetics* / Q000502:physiology; D011817:Rabbits; D051381:Rats; D016415:Sequence Alignment; D013552:Swine; D056189:Viral Tropism; D014779:Virus Replication / Q000502:physiology