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Selective flexible packaging pathways of the segmented genome of influenza A virus.

Nature communications (2020-08-30)
Ivan Haralampiev, Simon Prisner, Mor Nitzan, Matthias Schade, Fabian Jolmes, Max Schreiber, Maria Loidolt-Krüger, Kalle Jongen, Jasmine Chamiolo, Niklaas Nilson, Franziska Winter, Nir Friedman, Oliver Seitz, Thorsten Wolff, Andreas Herrmann
RESUMEN

The genome of influenza A viruses (IAV) is encoded in eight distinct viral ribonucleoproteins (vRNPs) that consist of negative sense viral RNA (vRNA) covered by the IAV nucleoprotein. Previous studies strongly support a selective packaging model by which vRNP segments are bundling to an octameric complex, which is integrated into budding virions. However, the pathway(s) generating a complete genome bundle is not known. We here use a multiplexed FISH assay to monitor all eight vRNAs in parallel in human lung epithelial cells. Analysis of 3.9 × 105 spots of colocalizing vRNAs provides quantitative insights into segment composition of vRNP complexes and, thus, implications for bundling routes. The complexes rarely contain multiple copies of a specific segment. The data suggest a selective packaging mechanism with limited flexibility by which vRNPs assemble into a complete IAV genome. We surmise that this flexibility forms an essential basis for the development of reassortant viruses with pandemic potential.

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Albumin, Acetylated from bovine serum, protease free, for molecular biology
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Lewatit® MonoPlus TP 214, ion exchange resin, macroporous