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

G.O. Acrani, N.L. Tilston-Lunel, M. Spiegel, M. Weidmann, M. Dilcher, D.E.A. Da Silva, M.R.T. Nunes, R.M. Elliott

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

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 39 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 cotransfecting a cDNA to the viral glycoproteins, virus-like particles were generated that packaged a minigenome and were capable of infecting naive cells.
Original languageEnglish
Pages (from-to)513-523
Number of pages11
JournalJournal of General Virology
Volume96
Issue number3
Early online date9 Dec 2014
DOIs
Publication statusPublished - Mar 2015

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