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Viruses 2016, 8(9), 247; doi:10.3390/v8090247

Binding of RNA by the Nucleoproteins of Influenza Viruses A and B

1
Institut de Biologie Structurale (IBS), University Grenoble Alpes, CEA, CNRS, 38044 Grenoble, France
2
EMBL Grenoble Outstation, 71 Avenue des Martyrs, BP181, F-38042 Grenoble CEDEX 9, France
*
Author to whom correspondence should be addressed.
Academic Editors: Roland Marquet and Polly Roy
Received: 8 July 2016 / Revised: 26 August 2016 / Accepted: 29 August 2016 / Published: 13 September 2016
(This article belongs to the Special Issue RNA Packaging)
View Full-Text   |   Download PDF [3240 KB, uploaded 13 September 2016]   |  

Abstract

This paper describes a biochemical study for making complexes between the nucleoprotein of influenza viruses A and B (A/NP and B/NP) and small RNAs (polyUC RNAs from 5 to 24 nucleotides (nt)), starting from monomeric proteins. We used negative stain electron microscopy, size exclusion chromatography-multi-angle laser light scattering (SEC-MALLS) analysis, and fluorescence anisotropy measurements to show how the NP-RNA complexes evolve. Both proteins make small oligomers with 24-nt RNAs, trimers for A/NP, and dimers, tetramers, and larger complexes for B/NP. With shorter RNAs, the affinities of NP are all in the same range at 50 mM NaCl, showing that the RNAs bind on the same site. The affinity of B/NP for a 24-nt RNA does not change with salt. However, the affinity of A/NP for a 24-nt RNA is lower at 150 and 300 mM NaCl, suggesting that the RNA binds to another site, either on the same protomer or on a neighbour protomer. For our fluorescence anisotropy experiments, we used 6-fluorescein amidite (FAM)-labelled RNAs. By using a (UC)6-FAM3′ RNA with 150 mM NaCl, we observed an interesting phenomenon that gives macromolecular complexes similar to the ribonucleoprotein particles purified from the viruses. View Full-Text
Keywords: influenza virus; nucleoprotein; RNA; oligomerization; assembly; ribonucleoprotein influenza virus; nucleoprotein; RNA; oligomerization; assembly; ribonucleoprotein
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MDPI and ACS Style

Labaronne, A.; Swale, C.; Monod, A.; Schoehn, G.; Crépin, T.; Ruigrok, R.W.H. Binding of RNA by the Nucleoproteins of Influenza Viruses A and B. Viruses 2016, 8, 247.

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