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

Acrani, G. O., Tilston-Lunel, N. L., Spiegel, M., Weidmann, M., Dilcher, M., Andrade da Silva, D.E., Nunes, M.R.T. and Elliott, R.M. (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(Pt_3), pp. 513-523. (doi: 10.1099/jgv.0.000005) (PMID:25491420)

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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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Elliott, Professor Richard
Authors: Acrani, G. O., Tilston-Lunel, N. L., Spiegel, M., Weidmann, M., Dilcher, M., Andrade da Silva, D.E., Nunes, M.R.T., and Elliott, R.M.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Journal Name:Journal of General Virology
Publisher:Society for General Microbiology
ISSN:0022-1317
ISSN (Online):1465-2099

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
635361Molecular analyses of arbovirus-host interactionsRichard ElliottWellcome Trust (WELLCOME)099220/B/12/ZMVLS III - CENTRE FOR VIRUS RESEARCH