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Import of adenovirus DNA involves the nuclear pore complex receptor CAN/Nup214 and histone H1

Abstract

Adenovirus type 2 (Ad2) imports its DNA genome through the nuclear pore complex (NPC) of cells in interphase for viral production. Here we identify the NPC-filament protein CAN/Nup214 as a docking site for incoming Ad2 capsids. Binding to CAN is independent of cytosolic factors. Capsids disassemble at NPCs to free their DNA for import. This process requires binding of nuclear histone H1 to the stably docked capsids and involves H1-import factors, restricting this irreversible process to the proximity of the nucleus. Our results provide a molecular mechanism for disassembly of Ad2 and reveal an unexpected function of histone H1 in virus-mediated DNA import.

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Figure 1: Ad2 binding to nuclear envelopes is cytosol- and Ran-independent.
Figure 2: The NPC protein CAN/Nup214 is a docking site for Ad2.
Figure 3: Antibody-targeting to CAN/Nup214 interferes with Ad2 disassembly in A549 cells.
Figure 4: Anti-CAN antibodies inhibit Ad2 import in vivo.
Figure 5: Histone H1.2 binds Ad2 in vitro and in vivo.
Figure 6: Ad2 but not Ad3 hexon contains a subgroup-specific acidic cluster.
Figure 7: Anti-histone H1 antibodies inhibit Ad2 disassembly and nuclear import of DNA in A549 cells.
Figure 8: Histone H1 and Ad2–TR colocalize after virus disassembly.
Figure 9: Model for Ad2 disassembly and DNA import.

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Acknowledgements

We thank P. Groscurth for access to the confocal laser scanning microscope and P. Sonderegger for access to the SMART system, U. Ziegler and P. Cinelli for assistance and U. Aebi, U. Kutay and T. Misteli for comments on the manuscript. Numerous investigators are gratefully acknowledged for their gifts of reagents (and are indicated in the Methods section) and H. van der Velde is acknowledged for preparation of C terminus CAN/Nup214. The work was supported by the Swiss National Science Foundation and the Kanton of Zürich.

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Figure S1 Microinjected anti-NPC antibodies reduce nuclear import of FITC–BSA–NLS. (PDF 614 kb)

Figure S2 Histone H1 import factors are required for nuclear import of Ad2–DNA.

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Trotman, L., Mosberger, N., Fornerod, M. et al. Import of adenovirus DNA involves the nuclear pore complex receptor CAN/Nup214 and histone H1. Nat Cell Biol 3, 1092–1100 (2001). https://doi.org/10.1038/ncb1201-1092

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