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Nat.Com. Nucleotide resolution mapping of influenza A virus nucleoprotein-RNA interactions reveals RNA features required for replication

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  • Nat.Com. Nucleotide resolution mapping of influenza A virus nucleoprotein-RNA interactions reveals RNA features required for replication

    Nat Commun. 2018 Jan 31;9(1):465. doi: 10.1038/s41467-018-02886-w.
    Nucleotide resolution mapping of influenza A virus nucleoprotein-RNA interactions reveals RNA features required for replication.

    Williams GD1,2, Townsend D2, Wylie KM3,4, Kim PJ5, Amarasinghe GK5,6, Kutluay SB2, Boon ACM7,8,9.
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    Abstract

    Influenza A virus nucleoprotein (NP) association with viral RNA (vRNA) is essential for packaging, but the pattern of NP binding to vRNA is unclear. Here we applied photoactivatable ribonucleoside enhanced cross-linking and immunoprecipitation (PAR-CLIP) to assess the native-state of NP-vRNA interactions in infected human cells. NP binds short fragments of RNA (~12 nucleotides) non-uniformly and without apparent sequence specificity. Moreover, NP binding is reduced at specific locations within the viral genome, including regions previously identified as required for viral genome segment packaging. Synonymous mutations designed to alter the predicted RNA structures in these low-NP-binding regions impact genome packaging and result in virus attenuation, whereas control mutations or mutagenesis of NP-bound regions have no effect. Finally, we demonstrate that the sequence conservation of low-NP-binding regions is required in multiple genome segments for propagation of diverse mammalian and avian IAV in host cells.


    PMID: 29386621 DOI: 10.1038/s41467-018-02886-w

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