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Journal Articles Viruses Year : 2020

Differential Behaviours and Preferential Bindings of Influenza Nucleoproteins on Importins-α

Abstract

Influenza viruses are negative single-stranded RNA viruses with nuclear transcription and replication. They enter the nucleus by using the cellular importin-α/-β nuclear import machinery. Influenza nucleoproteins from influenza A, B, C and D viruses possess a nuclear localization signal (NLS) localized on an intrinsically disordered extremity (NPTAIL). In this paper, using size exclusion chromatography (SEC), SEC-multi-angle laser light scattering (SEC-MALLS) analysis, surface plasmon resonance (SPR) and fluorescence anisotropy, we provide the first comparative study designed to dissect the interaction between the four NPTAILs and four importins-α identified as partners. All interactions between NPTAILs and importins-α have high association and dissociation rates and present a distinct and specific behaviour. D/NPTAIL interacts strongly with all importins-α while B/NPTAIL shows weak affinity for importins-α. A/NPTAIL and C/NPTAIL present preferential importin-α partners. Mutations in B/NPTAIL and D/NPTAIL show a loss of importin-α binding, confirming key NLS residues. Taken together, our results provide essential highlights of this complex translocation mechanism.
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Dates and versions

hal-02912888 , version 1 (23-11-2020)

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Amélie Donchet, Emilie Vassal-Stermann, Francine Gérard, Rob W H Ruigrok, Thibaut Crépin. Differential Behaviours and Preferential Bindings of Influenza Nucleoproteins on Importins-α. Viruses, 2020, 12 (8), pp.834. ⟨10.3390/v12080834⟩. ⟨hal-02912888⟩
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