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Published online
doi:10.1083/jcb.200707026
The Journal of Cell Biology, Vol. 180, No. 5, 857-865
The Rockefeller University Press, 0021-9525 $30.00
© Dultz et al.
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Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells



Elisa Dultz, Esther Zanin, Claudia Wurzenberger, Marion Braun, Gwénaël Rabut, Lucia Sironi, and Jan Ellenberg

Gene Expression Unit, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany

Correspondence to J. Ellenberg: jan.ellenberg{at}embl.de

During mitosis in higher eukaryotes, nuclear pore complexes (NPCs) disassemble in prophase and are rebuilt in anaphase and telophase. NPC formation is hypothesized to occur by the interaction of mitotically stable subcomplexes that form defined structural intermediates. To determine the sequence of events that lead to breakdown and reformation of functional NPCs during mitosis, we present here our quantitative assay based on confocal time-lapse microscopy of single dividing cells. We use this assay to systematically investigate the kinetics of dis- and reassembly for eight nucleoporin subcomplexes relative to nuclear transport in NRK cells, linking the assembly state of the NPC with its function. Our data establish that NPC assembly is an ordered stepwise process that leads to import function already in a partially assembled state. We furthermore find that nucleoporin dissociation does not occur in the reverse order from binding during assembly, which may indicate a distinct mechanism.

E. Dultz, E. Zanin, and C. Wurzenberger contributed equally to this paper.

Abbreviations used in this paper: IBB, importin β–binding domain of importin {alpha}; LBR, lamin B receptor; NE, nuclear envelope; NPC, nuclear pore complex; Nup, nucleoporin.


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