X-ray Structure of the Human Karyopherin RanBP5, an Essential Factor for Influenza Polymerase Nuclear Trafficking

https://doi.org/10.1016/j.jmb.2020.03.021Get rights and content

Highlights

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    RanBP5, by importing PA-PB1 polymerase sub-complex in the nucleus of the infected cells, is an essential factor of influenza virus infection.

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    The two isoform structures of human karyopherin RanBP5 illustrate its inherent dynamics.

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    We identify the NLS-binding site in RanBP5.

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    Using a binary protein complementation assay, we can suppress influenza PA-PB1 subcomplex interaction with RanBP5.

Abstract

Here, we describe the crystal structures of two distinct isoforms of ligand-free human karyopherin RanBP5 and investigate its global propensity to interact with influenza A virus polymerase. Our results confirm the general architecture and mechanism of the IMB3 karyopherin-β subfamily whilst also highlighting differences with the yeast orthologue Kap121p. Moreover, our results provide insight into the structural flexibility of β-importins in the unbound state. Based on docking of a nuclear localisation sequence, point mutations were designed, which suppress influenza PA-PB1 subcomplex binding to RanBP5 in a binary protein complementation assay.

Section snippets

Accession codes

Coordinates have been deposited to the PDB with IDs 6XTE (isoform-1) and 6XU2 (isoform-3).

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Supplementary material

Acknowledgments

We thank all members of our laboratories for their contributions and helpful discussions. C.S. was funded by the Hofmann-La Roche pRED external collaboration programme. This work was supported by the Agence Nationale pour la Recherche (ANR-14-CE09-0017, RNAP-IAV) and the European Commission (EC) framework program 7 project ComplexINC (contract no. 279039). This study used the platforms of the Grenoble Instruct Centre (ISBG; UMS 3518 CNRS-CEA-UJF-EMBL) with support from FRISBI (ANR-10-INSB-05-02

Declaration of Competing Interests

The authors declare no conflicts of interest.

Author contributions

Conceptualization: T.C.; Methodology: T.C., B.D. and C.B.; Investigation: C.S., B.D.C., L.S., F.G., A.A.M.C. and T.C.; Writing - Original Draft: C.S. and T.C.; Writing - Review & Editing: T.C., C.S., B.D., C.B., I.B. and R.W.H.R.; Supervision: T.C., B.D., C.B., I.B. and R.W.H.R.; Funding Acquisition: T.C., B.D., C.B., I.B. and R.W.H.R.

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    • †

    Present address: C. Swale, Institute for Advanced Biosciences (IAB), INSERM U1209, CNRS UMR 5309, University Grenoble Alpes, 38000 Grenoble, France.

    ‡

    Present address: F. Garzoni, Imophoron Ltd., UnitDX, St. Philips Central, Albert Road, Bristol BS2 OXJ, United Kingdom.

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