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.

Details

Title
Nucleotide resolution mapping of influenza A virus nucleoprotein-RNA interactions reveals RNA features required for replication
Author
Williams, Graham D 1 ; Townsend, Dana 2 ; Wylie, Kristine M 3 ; Kim, Preston J 4 ; Amarasinghe, Gaya K 5 ; Kutluay, Sebla B 2 ; Boon, Adrianus C M 6 

 Department of Medicine at Washington University School of Medicine, St Louis, MO, USA; Department of Molecular Microbiology at Washington University School of Medicine, St Louis, MO, USA 
 Department of Molecular Microbiology at Washington University School of Medicine, St Louis, MO, USA 
 Department of Pediatrics at Washington University School of Medicine, St Louis, MO, USA; The McDonnell Genome Institute at Washington University School of Medicine, St Louis, MO, USA 
 Department of Pathology and Immunology at Washington University School of Medicine, St Louis, MO, USA 
 Department of Pathology and Immunology at Washington University School of Medicine, St Louis, MO, USA; Department of Biochemistry and Biophysics at Washington University School of Medicine, St Louis, MO, USA 
 Department of Medicine at Washington University School of Medicine, St Louis, MO, USA; Department of Molecular Microbiology at Washington University School of Medicine, St Louis, MO, USA; Department of Pathology and Immunology at Washington University School of Medicine, St Louis, MO, USA 
Pages
1-12
Publication year
2018
Publication date
Jan 2018
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
1992989549
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.