Binding of RNA by the Nucleoproteins of Influenza Viruses A and B
AbstractThis 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
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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.
Labaronne A, Swale C, Monod A, Schoehn G, Crépin T, Ruigrok RWH. Binding of RNA by the Nucleoproteins of Influenza Viruses A and B. Viruses. 2016; 8(9):247.Chicago/Turabian Style
Labaronne, Alice; Swale, Christopher; Monod, Alexandre; Schoehn, Guy; Crépin, Thibaut; Ruigrok, Rob W.H. 2016. "Binding of RNA by the Nucleoproteins of Influenza Viruses A and B." Viruses 8, no. 9: 247.
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