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

Citation

Acrani GO, Tilston-Lunel NL, Spiegel M, Weidmann M, Dilcher M, Silva DEA, Nunes MRT & Elliott RM (2015) Establishment of a minigenome system for oropouche virus reveals the S genome segment to be significantly longer than reported previously. Journal of General Virology, 96 (3), pp. 513-523. https://doi.org/10.1099/jgv.0.000005

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.

Journal
Journal of General Virology: Volume 96, Issue 3

• Acrani et al_JGV_2015.pdf