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Dultz, E. et al. Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells. J. Cell Biol. 180, 857-865

Gene Expression Unit, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany.
The Journal of Cell Biology (Impact Factor: 9.69). 04/2008; 180(5):857-65. DOI: 10.1083/jcb.200707026
Source: PubMed

ABSTRACT 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.

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    • "Nup93 subsequently recruits the FG repeat-containing nucleoporins of the Nup62 complex. The FG-containing nucleoporin Nup98 is recruited concomitantly with Nup93 (Dultz et al. 2008) and has recently been found to be key to the establishment of the transport and exclusion properties of the pore (Hulsmann et al. 2012; Laurell et al. 2011). Together, these FG nucleoporins form a substantial part of the hydrophobic meshwork in the center of the pore (Ribbeck and Gorlich 2001). "
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    • "Nup93 subsequently recruits the FG repeat-containing nucleoporins of the Nup62 complex. The FG-containing nucleoporin Nup98 is recruited concomitantly with Nup93 (Dultz et al. 2008) and has recently been found to be key to the establishment of the transport and exclusion properties of the pore (Hulsmann et al. 2012; Laurell et al. 2011). Together, these FG nucleoporins form a substantial part of the hydrophobic meshwork in the center of the pore (Ribbeck and Gorlich 2001). "
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    ABSTRACT: The metazoan nucleus is disassembled and re-built at every mitotic cell division. The nuclear envelope, including nuclear pore complexes, breaks down at the beginning of mitosis to accommodate the capture of massively condensed chromosomes by the spindle apparatus. At the end of mitosis, a nuclear envelope is newly formed around each set of segregating and de-condensing chromatin. We review the current understanding of the membrane restructuring events involved in the formation of the nuclear membrane sheets of the envelope, the mechanisms governing nuclear pore complex assembly and integration in the nascent nuclear membranes, and the regulated coordination of these events with chromatin de-condensation.
    Chromosoma 10/2012; 121(6). DOI:10.1007/s00412-012-0388-3 · 3.26 Impact Factor
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    • "Similarly, C. elegans Nup53 is necessary for the efficient accumulation of Nup155 and Nup58 but not Nup107 at the NE (Rodenas et al, 2009). This is also supported by live-cell imaging experiments in HeLa cells, which capture the recruitment of Nup58 slightly after Nup93 (Dultz et al, 2008). Accordingly, we have found that upon depletion of the two Nup93 containing subcomplexes, Nup93–Nup188 and Nup93–205, the two other members of the complex, Nup155 and Nup53, are still detectable, albeit at reduced levels on the assembling NPCs (Sachdev et al, 2012). "
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    ABSTRACT: Nuclear pore complexes (NPCs) fuse the two membranes of the nuclear envelope (NE) to a pore, connecting cytoplasm and nucleoplasm and allowing exchange of macromolecules between these compartments. Most NPC proteins do not contain integral membrane domains and thus it is largely unclear how NPCs are embedded and anchored in the NE. Here, we show that the evolutionary conserved nuclear pore protein Nup53 binds independently of other proteins to membranes, a property that is crucial for NPC assembly and conserved between yeast and vertebrates. The vertebrate protein comprises two membrane binding sites, of which the C-terminal domain has membrane deforming capabilities, and is specifically required for de novo NPC assembly and insertion into the intact NE during interphase. Dimerization of Nup53 contributes to its membrane interaction and is crucial for its function in NPC assembly.
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