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Development of an Isavirus minigenome system to study the function of the pocket RNA-binding domain of the viral nucleoprotein (NP) in salmon cells

dc.contributor.authorToro-Ascuy D.
dc.contributor.authorSantibañez A.
dc.contributor.authorPeña V.
dc.contributor.authorBeltran-Pavez C.
dc.contributor.authorCottet L.
dc.contributor.authorMolina C.
dc.contributor.authorMontoya M.
dc.contributor.authorSandoval N.
dc.contributor.authorVásquez-Martínez Y.
dc.contributor.authorMascayano C.
dc.contributor.authorCortez-San Martín M.
dc.date.accessioned2020-09-02T22:29:17Z
dc.date.available2020-09-02T22:29:17Z
dc.date.issued2020
dc.identifier10.1111/jfd.13122
dc.identifier.citation43, 2, 197-206
dc.identifier.issn01407775
dc.identifier.urihttps://hdl.handle.net/20.500.12728/6402
dc.descriptionThe Isavirus is an orthomyxovirus with a genome composed of eight segments of negative single-strand RNA (−ssRNA). It has been proposed that the eight genomic segments of the Isavirus are organized as a ribonucleoprotein (RNP) complex called a minigenome, which contains all the viral RNA segments, a viral heterotrimeric polymerase and multiple copies of the viral nucleoprotein (NP). Here, we develop an Isavirus minigenome system and show the importance of the formation of active RNPs and the role of viral NP R189, R194, R302 and K325 residues in the NP RNA-binding domain in the context of RNPs. The results indicate it is possible to generate a minigenome in salmon cells, a composite ISAV RNPs with EGFP-based chimeric vRNA with heterotrimeric polymerase (PB1, PB2, PA) and NP protein using CMV-based auxiliary plasmids. It was also shown that NP R189, R194, R302 and K325 residues are important to generate viral mRNA from the constituted RNPs and a detectable reporter protein. This work is the first salmon cell-based minigenome assay for the Isavirus, which was evaluated by a bioinformatic and functional study of the NP protein in viral RNPs, which showed that correct NP-vRNA interaction is key to the functioning of RNPs. © 2019 John Wiley & Sons Ltd
dc.language.isoen
dc.publisherBlackwell Publishing Ltd
dc.subjectinfectious salmon anaemia virus
dc.subjectminigenome assay
dc.subjectnucleoprotein
dc.subjectorthomyxovirus
dc.subjectsalmon
dc.subjectmessenger RNA
dc.subjectvirus nucleoprotein
dc.subjectribonucleoprotein
dc.subjectviral protein
dc.subjectanimal cell
dc.subjectArticle
dc.subjectassay
dc.subjectIsavirus
dc.subjectnonhuman
dc.subjectplasmid
dc.subjectRNA-binding domain
dc.subjectsalmonine
dc.subjectvirus genome
dc.subjectanimal
dc.subjectgenetics
dc.subjectgenomics
dc.subjectIsavirus
dc.subjectRNA-binding domain
dc.subjectSalmo salar
dc.subjectvirology
dc.subjectAnimals
dc.subjectGenome, Viral
dc.subjectGenomics
dc.subjectIsavirus
dc.subjectRibonucleoproteins
dc.subjectRNA-Binding Motifs
dc.subjectSalmo salar
dc.subjectViral Proteins
dc.titleDevelopment of an Isavirus minigenome system to study the function of the pocket RNA-binding domain of the viral nucleoprotein (NP) in salmon cells
dc.typeArticle


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