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RanGTP mediates nuclear pore complex assembly

Nature volume 424, pages 689–694 (2003)Cite this article

Abstract

In metazoa, the nuclear envelope breaks down and reforms during each cell cycle. Nuclear pore complexes (NPCs), which serve as channels for transport between the nucleus and cytoplasm1, assemble into the reforming nuclear envelope in a sequential process involving association of a subset of NPC proteins, nucleoporins, with chromatin followed by the formation of a closed nuclear envelope fenestrated by NPCs2,3,4,5,6,7. How chromatin recruitment of nucleoporins and NPC assembly are regulated is unknown. Here we demonstrate that RanGTP production is required to dissociate nucleoporins Nup107, Nup153 and Nup358 from Importin β, to target them to chromatin and to induce association between separate NPC subcomplexes. Additionally, either an excess of RanGTP or removal of Importin β induces formation of NPC-containing membrane structures—annulate lamellae—both in vitro in the absence of chromatin and in vivo. Annulate lamellae formation is strongly and specifically inhibited by an excess of Importin β. The data demonstrate that RanGTP triggers distinct steps of NPC assembly, and suggest a mechanism for the spatial restriction of NPC assembly to the surface of chromatin.

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Figure 1: RanGTP stimulates nucleoporin recruitment to sperm chromatin.
Figure 2: Ran GTP is required for post-mitotic AL assembly.
Figure 3: RanGTP-mediated release of Importin β from nucleoporin complexes.
Figure 4: Depletion of Importin β induces the assembly of nucleoporins into membranes.

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Acknowledgements

We thank C. Schatz for recombinant RanBP5, V. Galy for experimental advice, G. Krohne for antibodies, K. Ribbeck for discussions and W. Antonin, J. Ellenberg, V. Galy, O. Gruss and E. Izaurralde for critically reading the manuscript. M.H. was supported by the Austrian Programme for Advanced Research and Technology.

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  1. European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117, Heidelberg, Germany

    Tobias C. Walther, Peter Askjaer, Marc Gentzel, Anja Habermann, Gareth Griffiths, Matthias Wilm, Iain W. Mattaj & Martin Hetzer

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Correspondence to Iain W. Mattaj or Martin Hetzer.

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Walther, T., Askjaer, P., Gentzel, M. et al. RanGTP mediates nuclear pore complex assembly. Nature 424, 689–694 (2003). https://doi.org/10.1038/nature01898

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