title: Establishment of a minigenome system for Oropouche virus reveals the S genome segment to be significantly longer than reported previously.
abstract: 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.
authors.affiliation: Department of Cell and Molecular Biology, University of Sao Paulo School of Medicine, 3900, Av. Bandeirantes, Ribeirão Preto, SP 14049-900, Brazil.|MRC-University of Glasgow Centre for Virus Research, 464 Bearsden Road, Glasgow G61 1QH, Scotland, UK.
authors.forename: Gustavo Olszanski
authors.initials: GO
authors.lastname: Acrani
authors.affiliation: Biomedical Sciences Research Complex, School of Biology, University of St Andrews, St Andrews KY16 9ST, Scotland, UK.|MRC-University of Glasgow Centre for Virus Research, 464 Bearsden Road, Glasgow G61 1QH, Scotland, UK.
authors.forename: Natasha L
authors.initials: NL
authors.lastname: Tilston-Lunel
authors.affiliation: Department of Virology, University Medical Center Göttingen, Kreuzbergring 57, 37075 Göttingen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Spiegel
authors.affiliation: Department of Virology, University Medical Center Göttingen, Kreuzbergring 57, 37075 Göttingen, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Weidmann
authors.affiliation: Department of Virology, University Medical Center Göttingen, Kreuzbergring 57, 37075 Göttingen, Germany.
authors.forename: Meik
authors.initials: M
authors.lastname: Dilcher
authors.affiliation: Center for Technological Innovation, Instituto Evandro Chagas, Ananindeua, Brazil.
authors.forename: Daisy Elaine
authors.initials: DE
authors.lastname: Andrade da Silva
authors.affiliation: Center for Technological Innovation, Instituto Evandro Chagas, Ananindeua, Brazil.
authors.forename: Marcio R T
authors.initials: MRT
authors.lastname: Nunes
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, 464 Bearsden Road, Glasgow G61 1QH, Scotland, UK.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Elliott
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D016679:Genome, Viral / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D016851:Orthobunyavirus / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D017423:Sequence Analysis, RNA
chemical_list: D012367:RNA, Viral