Establishment of a minigenome system for Oropouche virus reveals the S genome segment to be significantly longer than reported previously

• Autor(a) principal: Acrani, Gustavo Olszanski
• Data de Publicação: 2015
• Outros Autores: Tilston-Lunel, Natasha L, Spiegel, Martin, Weidmann, Manfred, Dilcher, Meik, Silva, Daisy Elaine Andrade da, Nunes, Márcio Roberto Teixeira, Elliott, Richard M
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Digital do Instituto Evandro Chagas (Patuá)
• Texto Completo: https://patua.iec.gov.br/handle/iec/3858
• topic: Orthobunyavirus / genética; Genoma Viral; Clonagem Molecular; Genômica / métodos; Dados de Sequência Molecular; RNA Viral / genética

Oropouche virus (OROV) is a medically important orthobunyavirus, which causes frequent outbreaks of a febrile illness in the northern parts of Brazil. However, despite being the cause of an estimated half a million human infections since its first isolation in Trinidad in 1955, details of the molecular biology of this tripartite, negative-sense RNA virus remain limited. We have determined the complete nucleotide sequence of the Brazilian prototype strain of OROV, BeAn 19991, and found a number of differences compared with sequences in the database. Most notable were that the S segment contained an additional 204 nt at the 3' end and that there was a critical nucleotide mismatch at position 9 within the base-paired terminal panhandle structure of each genome segment. In addition, we obtained the complete sequence of the Trinidadian prototype strain TRVL-9760 that showed similar characteristics to the BeAn 19991 strain. By using a T7 RNA polymerase-driven minigenome system, we demonstrated that cDNA clones of the BeAn 19991 L and S segments expressed functional proteins, and also that the newly determined terminal untranslated sequences acted as functional promoters in the minigenome assay. By co-transfecting a cDNA to the viral glycoproteins, virus-like particles were generated that packaged a minigenome and were capable of infecting naive cells.