Published August 8, 2011

Text by Ben Short bshort@rockefeller.edu

In Focus

Nuclear envelope star ts with a clean sheet

After mitosis, nuclear membranes form directly from ER cisternae before nuclear pore complex reassembly.
t the start of mitosis, the nuclear FOCAL POINT envelope breaks down and merges with the endoplasmic reticulum (ER). At the same time, the nuclear pore complexes that mediate transport across the inner and outer membranes of the envelope also disassemble. Transmembrane pore proteins move into the ER, while soluble components disperse into the cytoplasm. Lu et al. use rapid, live-cell 3D imaging to establish the sequence of events as the nuclear envelope and pores (Left to right) Lei Lu, Mark Ladinsky, and Tomas Kirchhausen describe the sequence of events by which the nuclear envelope and nuclear pore complexes reassemble after mitosis. In contrast to are rebuilt after mitosis (1). previous models, Lu et al. nd that the nuclear envelope forms directly from mitotic ER cisternae. The nuclear envelope has been proposed A 3D model constructed by electron tomography (far right) shows an ER sheet (green) contacting to reassemble through the fusion of ER the surface of mitotic chromosomes (blue) and splitting into the inner and outer membranes of tubules around the chromosomes (2, 3). the nascent nuclear envelope. The researchers also demonstrate that nuclear pore complexes In 2009, however, Tomas Kirchhausen and only form after nuclear envelope reassembly, rather than preassembling on chromatin in the his postdoc Lei Lu from Harvard Medical absence of nuclear membranes. School in Boston found that the ER mainly exists as large, sheet-like cisternae during In support of this, the researchers found masses already covered by nuclear envemitosis (4). This suggested to us that these that depolymerizing spindle microtubules lope. Moreover, by calibrating and quancisternae provide the membrane for nuclear with nocodazole increased the rate of nu- tifying the fluorescence of their nuclear envelope reformation, says Lu, who is now clear membrane reassembly, whereas sta- pore marker, the researchers determined starting his own laboratory at Nanyang bilizing the spindle with taxol delayed that the small amount of pore protein to Technological University in Singapore. envelope reformation. Because the dynam- accumulate on chromosomes before nuclear Lu and Kirchhausen therefore reexam- ics of spindle disassembly vary in different envelope formation corresponded to single ined nuclear envelope reassembly in live cell types, nuclear envelope formation protein units rather than higher-order preHeLa cells expressing fluorescent markers may differ in cell lines other than HeLa pore structures. of the ER and chromatin Lu et al. saw cisternae iniNuclear pores are therefore always as(1). The researchers found tiate envelope assembly all sembled within a preexisting nuclear enveIt unifies what over the anaphase chromo- lope. It unifies what happens in interphase, that, during anaphase, ER sheets first contacted the some masses of BSC1 cells, what happens in mitosis, and what happens happens in rims of the disk-shaped example. interphase [with] for The researchers then in yeast [which undergo closed mitoses], chromosome masses movsays Kirchhausen. Of course there will be what happens ing to opposite spindle poles turned their attention to the differences but, topologically, you dont before expanding to fully assembly of nuclear pores have to postulate different models. in mitosis. enclose the daughter cell after mitosis. During interLu and Kirchhausen now want to innuclei. In collaboration with phase, nuclear pore compo- vestigate the details of how nuclear pores Mark Ladinsky from Caltech in Pasadena, nents form new pore complexes by inserting reassemble after mitosis. We want to genLu and Kirchhausen used high-resolution themselves into the nuclear envelope. The erate a clearer molecular picture of how electron tomography to confirm that the ER same mechanism might apply to post- nuclear pore complexes make the hole in membranes enveloping the chromosomes mitotic pore assembly, but some groups the intact nuclear membranes, Lu says. were cisternal, rather than tubular, in shape. have proposed an alternative pathway in 1. Lu, L., et al. 2011. J. Cell Biol. doi:10.1083/ Lu et al. think that ER cisternae might which pore proteins form prepore comjcb.201012063. initially target the rim of chromosome plexes on the surface of the chromosome 2. Anderson, D.J., and M.W. Hetzer. 2007. Nat. Cell masses in HeLa cells because the mitotic masses in early anaphase before nuclear Biol. 9:11601166. spindle blocks the ER from contacting the envelope reformation (5, 6). 3. Anderson, D.J., and M.W. Hetzer. 2008. J. Cell Biol. 182:911924. outer and inner faces of the chromatin By imaging live cells expressing fluoresdisks. We dont think theres any special cent markers of the ER and nuclear pores, 4. Lu, L., et al. 2009. Mol. Biol. Cell. 20:34713480. signal, explains Kirchhausen. [The rim] is Lu et al. found that pore complexes only 5. Walther, T.C., et al. 2003. Cell. 113:195206. simply where you have topological access. reassembled in regions of the chromosome 6. Dultz, E., et al. 2008. J. Cell Biol. 180:857865.

LU PHOTO COURTESY OF TIE HIENG CHONG; LADINSKY PHOTO COURTESY OF ARIANE BRIEGEL, KIRCHHAUSEN PHOTO COURTESY OF THE AUTHOR

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