Fichtman B , Ramos C , Rasala B , Harel A , Forbes DJ .

R01 GM-33279 NIGMS NIH HHS , R01 GM033279-27 NIGMS NIH HHS , R01 GM033279-28 NIGMS NIH HHS , R01 GM033279 NIGMS NIH HHS

	Figure 1. Cold temperature produces a novel intermediate in NPC assembly at 30 min that lacks FG Nups. (A) The progression of nuclear envelope assembly is slowed down at 14°C. Nuclear assembly reactions containing chromatin, cytosol, and membranes were set up at different temperatures. Reactions were stopped at the indicated time points and analyzed by confocal microscopy after direct immunofluorescence with the Alexa568-labeled anti-FG Nup antibody mAb414 (red). Nuclear membranes were visualized with the fluorescent membrane dye DHCC (green), and chromatin was stained with DAPI (blue). Scale bar, 10 μm. (B) A DNA antibody exclusion assay was used to determine the formation of closed nuclear envelopes (CNE). For this, nuclear assembly reactions containing chromatin, cytosol and membranes were set up and incubated for 30 min at room temperature (RT), for 30 min at 14°C, and for 30 min at RT in the presence of BAPTA before fixation. Alexa568-labeled anti-DNA antibody (red) and Alexa488-labeled anti-FG Nup antibody (green) were added and the nuclei examined. Chromatin plus cytosol (no membranes) served as a positive control for anti-DNA antibody binding (top panel). Scale bar, 10 μm. (C) The formation of CNE was quantified for at least 100 randomly chosen structures in three independent experiments. (D) The percentage of CNE containing FG Nups, detected as a green rim with Alexa488-labeled anti-FG Nup antibody at 30 min and 60 min in three different experiments was also plotted. Errors bars indicate the SD. Nuclei reconstituted in equivalent conditions of 30 min 14°C are marked with a diamond for anti-DNA antibody (♦) and an asterisk (*) for anti-FG Nup antibody in panels A–D.
	Figure 2. The 30-min cold intermediate contains punctate POM121 and Nup133 on the inner face of the nuclear envelope. (A) A nuclear assembly reaction was incubated for 30 min at 14°C and analyzed by confocal microscopy after direct immunofluorescence on ice with Alexa568-anti-POM121 (red) and Alexa488-anti-Nup133 (green) antibodies (left panels). The staining of the same nuclear intermediates with Alexa647-anti-DNA antibody (gray) is shown in the right panels; this latter staining was used to assess whether an individual nucleus contained a fully closed (top panels) or still open (bottom panels) nuclear envelope. Merged images of POM121/Nup133/DAPI stain are shown at normal (left panels) and high (adjacent panels) magnification. We note that POM121 and Nup133 could only be stained in intermediates where the nuclear envelope was open (i.e., accessible to anti-DNA antibody). A contour trace of the POM121 and Nup133 staining is shown in the 3rd set of panels, allowing comparison of their localization. Scale bar, 10 μm. (B) Nuclei with accessible interiors were analyzed by direct immunofluorescence with Alexa568-anti-POM121 (red) and Alexa488-anti-FG Nup (green) antibodies. The merged images (yellow) are shown at the right. Chromatin was stained with DAPI (blue). Scale bar, 10 μm. Individual NPCs were separately visualized at high magnification on the surface of 14°C nuclear intermediates assembled for 30 min and 60 min (surface). Scale bar, 1 μm. (C) The assembly reactions were analyzed in parallel with Alexa568-anti-Nup133 (red) and Alexa488-anti-FG Nup (green) antibodies. Scale bar, 10 μm.
	Figure 3. The 30-min cold intermediate containing POM121 and the Nup107-160 complex lacks diffusion channels. (A) An anti-DNA antibody exclusion assay was used to assess the formation of completely enclosed nuclear envelopes. Nuclei were assembled at room temperature for 60 min with chromatin, membranes, and either control cytosol, GTPγS-treated cytosol, or BAPTA-treated cytosol. Control nuclei were either visualized alone (Ctrl) or permeabilized with Triton X-100 (TX100) to allow entry of anti-DNA antibody. Positive labeling of the chromatin with Alexa568-anti-DNA antibody in the GTPγS nuclei indicated that a fully closed NE did not form under this condition; absence of anti-DNA antibody staining for Control and BAPTA nuclei indicated that a fully closed NE formed in these conditions. Scale bar, 10 μm. The percentage of nuclei showing a lack of DNA antibody staining in their nuclear interior (% CNE) was quantified for ∼100 randomly chosen nuclear intermediates each in three independent experiments, as shown in the histogram. Error bars indicate SD. (B) The anti-Alexa488 quenching assay described in Supplemental Figure 4 was used to determine the presence of 10-kDa competent diffusion channels in closed nuclear envelopes reconstituted for 15 min and 30 min at 14°C. The 15 min, 14°C intermediates were not fully membrane enclosed, as they showed anti-DNA staining (red) and quenching by anti-Alexa488 antibody (black) (top panels). By 30 min at 14°C, many intact nuclei had formed, as judged by a lack of DNA staining, but these showed no entry of Alexa488-10 kDa dextran indicating no diffusion channels (middle panels). In contrast, by 30 min at room temperature, the nuclei were intact (no anti-DNA staining) but had developed nuclear pores capable of allowing Alexa488-dextran entry, enabling its protection from the quenching antibody (lower panels). (C) The percentage of 14°C 30 min enclosed nuclear intermediates (i.e., with no anti-DNA staining) that showed diffusion of Alexa488-10 kDa or Alexa488-3 kDa dextrans was measured in three different experiments each and plotted. We found that the great majority of enclosed 14°C 30 min nuclear intermediates did not show diffusion channels. Errors bars indicate the SD.
	Figure 4. Channel formation is a late event in NPC assembly. To ask whether the nuclear intermediates that did not show diffusion channels by 30 min at 14°C were competent to develop such channels, the diffusion assay in Figure 3B was performed for both 30 min and 60 min at 14°C. Only nuclei showing closed nuclear envelopes (CNE) according to the anti-DNA antibody staining assay were analyzed for 3-kDa dextran uptake and were quantitated. At 30 min at 14°C, only 4% of intact nuclear intermediates showed internal Alexa488-3 kDa dextran. By 60 min at 14°C, however, 100% of the intact nuclear intermediates showed internal protected Alexa488-3 kDa dextran. Scale bars, 10 μm.
	Figure 5. LPC blocks diffusion channel formation. Nuclei were assembled for 30 min at 14°C, and the preparation was verified for the absence of diffusion channels, as in Figure 3B, with Alexa488-10 kDa dextran (left panels). The reaction was then supplemented with buffer, LPC, or LPC plus OA, followed by incubation for 90 min at 14°C to assess the affect of LPC on channel formation. The nuclei were assayed for channel formation with Alexa488-10 kDa dextran. A block to channel formation was observed with LPC (asterisk). When LPC and OA were added together, however, diffusion of Alexa488-10 kDa dextran into the nuclei was now observed. Control nuclei assembled at room temperature for 60 min that were the treated with Triton X-100 before the addition of antibodies served as a positive control for the anti-DNA antibody assay and for the dextran quenching assay (bottom panels). Scale bar, 10 μm.
	Figure 6. The cold intermediate contains all the membrane proteins required for NPC assembly. To determine whether the 14°C 30-min cold intermediate is able to go on to form nuclear pores without the addition of extra membranes, WTαSNAP was added as a vesicle–vesicle fusion inhibitor. Δ80NαSNAP served as a negative control. (A) We first looked at BAPTA-arrested pore-free nuclei which had been formed at room temperature for 30 min (left panels). Because neither SNAP protein is membrane permeable, we expected that WTαSNAP should inhibit addition of new vesicles to the forming nuclei, but could not inhibit inner-outer nuclear membrane fusion, thus permitting nuclear pore formation. Consistent with this, 30-min BAPTA-arrested pore-free nuclei which had been formed at room temperature, when diluted 1:10 into cytosol supplemented with buffer, WTαSNAP, or Δ80NαSNAP and incubated for a further 30 min, all became FG Nup–positive, as detected by Alexa488-anti-FG Nup antibody (green). (B) We performed the same type of experiment with nuclei reconstituted at 14°C for 30 min, at which time the reaction contained ∼50% closed, FG-negative nuclear intermediates. (Because of SNAP inhibition of vesicle–vesicle fusion, the open intermediates did not progress further.) The 14°C 30 min nuclei were diluted 1:10 into cytosol containing buffer, WTαSNAP (47 μM), or Δ80NαSNAP (47 μM) and incubated for a further 30 min at 14°C. Reactions were stopped on ice, fixed, and incubated on ice with Alexa488-anti-DNA (green) and Alexa568-anti-FG Nup (red) antibodies. All three reactions produced abundant anti-DNA–negative, FG-Nup–positive nuceli (red). Nuclear intermediates which had been assembled for 15 min at 14°C served as a negative control for the anti-FG Nup antibody (0%) and a positive control for the anti-DNA antibody (100%). A representative nucleus is shown in the bottom panels. The FG Nup–positive nuclei in B were quantified for each of three replicate experiments and the average percentage containing FG Nups is plotted in C. Errors bars indicate the SD.
	Figure 7. The fusion inhibitors LPC and OA individually prevent FG Nup assembly in the 30 min cold intermediate. To ask whether membrane fusion inhibitors prevent formation of mature nuclear pores in the 30-min cold intermediate, intermediates were first formed at 14°C for 30 min and verified with Alexa647-anti-DNA and Alexa488-anti-FG Nup antibodies. Approximately 95% of the nuclei showed no FG Nup staining (B, left column). (A) At this point LPC, OA (final concentration 600 μM each), or a mix of both were added and the reactions were incubated at 14°C for an additional 90 min. The reactions were stopped on ice, fixed with 2.5% paraformaldehyde for 30 min on ice, followed by incubation for 30 min on ice with Alexa488-anti-FG Nup antibody (green). LPC or OA blocked FG recruitment (asterisks). (B) The data in A are quantitated, and error bars indicate SD. (C) In parallel, an aliquot of the nuclei in the fixed reactions from A were stained with a mixture of Alexa568-anti-POM121 (red), Alexa488-anti-Nup133 (green), and Alexa674-anti-DNA antibodies (gray). In A, LPC- or OA-treated nuclei at 120-min total incubation showed little FG Nup staining (A and B, asterisks), indicating no progression to an FG Nup–positive state. However, when LPC and OA were added together (A, B, and C), nuclear assembly occurred normally in that the majority of the 120 min closed nuclei showed POM121, Nup133, and FG Nup staining (LPC+OA).
	Figure 8. A schematic model for inner/outer nuclear membrane fusion in the early steps of vertebrate NPC assembly. In previous studies, ELYS was shown to mediate the binding of the Nup107-160 complex to sites of AT-rich chromatin (blue) in early nuclear assembly. Simultaneous with this ER-derived membranes are recruited to the rim of developing nuclei. The data presented here argues that next: (1) the Nup107-160 complex (detected by anti-Nup133 antibody) interacts with POM121 at the inner nuclear membrane in an early nuclear intermediate (30 min, 14°C). Bending of the membranes toward one another would theoretically occur at or near this time. (2) This is followed by fusion of the inner and outer nuclear membranes to form a diffusion channel. (3) This fusion is simultaneous with or precedes recruitment of the bulk of FG nucleoporins (gold) and assembly of the mature nuclear pore, which involves expansion of the initial channel and its stabilization. In pioneering membrane-membrane fusion studies, progression to hemifusion was seen to be typically inhibited by cone-shaped lipids, such as LPC; here LPC was found to indeed block channel formation and FG Nup assembly in nuclear intermediates. Progression beyond hemifusion to full fusion has been observed to be inhibited by inverted cone-shaped lipids, such as OA; here we found that OA also blocks mature nuclear pore assembly. When added together LPC and OA are known to geometrically neutralize one another, and we observed that LPC+OA allow channel formation, i.e., inner/outer membrane fusion. (It is important to note that we do not believe that a completely sealed nuclear envelope is required before nuclear pores can form, but possession of such a sealed intermediate allowed us to distinguish and analyze the channel formation step of nuclear pore assembly.)

Anderson, Nuclear envelope formation by chromatin-mediated reorganization of the endoplasmic reticulum. 2007, Pubmed, Xenbase

Anderson, Nuclear envelope formation by chromatin-mediated reorganization of the endoplasmic reticulum. 2007, Pubmed , Xenbase
Antonin, Nuclear pore complex assembly through the cell cycle: regulation and membrane organization. 2008, Pubmed
Antonin, The integral membrane nucleoporin pom121 functionally links nuclear pore complex assembly and nuclear envelope formation. 2005, Pubmed , Xenbase
Baur, NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts. 2007, Pubmed , Xenbase
Beck, Nuclear pore complex structure and dynamics revealed by cryoelectron tomography. 2004, Pubmed
Belgareh, An evolutionarily conserved NPC subcomplex, which redistributes in part to kinetochores in mammalian cells. 2001, Pubmed
Bishop, Assembly of the endoplasmic reticulum phospholipid bilayer: the phosphatidylcholine transporter. 1985, Pubmed
Blobel, A potential fusion peptide and an integrin ligand domain in a protein active in sperm-egg fusion. 1992, Pubmed
Blower, A Rae1-containing ribonucleoprotein complex is required for mitotic spindle assembly. 2005, Pubmed , Xenbase
Boehmer, Purification, crystallization and preliminary X-ray analysis of a Nup107-Nup133 heterodimeric nucleoporin complex. 2007, Pubmed
Boman, GTP hydrolysis is required for vesicle fusion during nuclear envelope assembly in vitro. 1992, Pubmed , Xenbase
Brohawn, Molecular architecture of the Nup84-Nup145C-Sec13 edge element in the nuclear pore complex lattice. 2009, Pubmed
Burke, Nuclei take a position: managing nuclear location. 2009, Pubmed
Buton, Ultrafast glycerophospholipid-selective transbilayer motion mediated by a protein in the endoplasmic reticulum membrane. 1996, Pubmed
Chakraborty, Nucleoporin levels regulate cell cycle progression and phase-specific gene expression. 2008, Pubmed
Chernomordik, Membranes of the world unite! 2006, Pubmed
Chernomordik, The pathway of membrane fusion catalyzed by influenza hemagglutinin: restriction of lipids, hemifusion, and lipidic fusion pore formation. 1998, Pubmed
Chernomordik, Protein-lipid interplay in fusion and fission of biological membranes. 2003, Pubmed
Chernomordik, An early stage of membrane fusion mediated by the low pH conformation of influenza hemagglutinin depends upon membrane lipids. 1997, Pubmed
Chernomordik, Biomembrane fusion: a new concept derived from model studies using two interacting planar lipid bilayers. 1987, Pubmed
Chernomordik, Membrane hemifusion: crossing a chasm in two leaps. 2005, Pubmed
Chernomordik, Lysolipids reversibly inhibit Ca(2+)-, GTP- and pH-dependent fusion of biological membranes. 1993, Pubmed
Cilia, Plasmodesmata form and function. 2004, Pubmed
Cohen, Transmission electron microscope studies of the nuclear envelope in Caenorhabditis elegans embryos. 2002, Pubmed
Cotter, Nuclear pore complex disassembly and nuclear envelope breakdown during mitosis may occur by both nuclear envelope vesicularisation and dispersion throughout the endoplasmic reticulum. 1998, Pubmed , Xenbase
Crisp, The nuclear envelope as an integrator of nuclear and cytoplasmic architecture. 2008, Pubmed
D'Angelo, Structure, dynamics and function of nuclear pore complexes. 2008, Pubmed
D'Angelo, Nuclear pores form de novo from both sides of the nuclear envelope. 2006, Pubmed , Xenbase
Davuluri, Mutation of the zebrafish nucleoporin elys sensitizes tissue progenitors to replication stress. 2008, Pubmed , Xenbase
Dawson, ER membrane-bending proteins are necessary for de novo nuclear pore formation. 2009, Pubmed , Xenbase
Debler, A fence-like coat for the nuclear pore membrane. 2008, Pubmed
de Bruyn Kops, An essential nuclear envelope integral membrane protein, Brr6p, required for nuclear transport. 2001, Pubmed
Dechat, Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin. 2008, Pubmed
DeGrasse, Evidence for a shared nuclear pore complex architecture that is conserved from the last common eukaryotic ancestor. 2009, Pubmed
de Jong-Curtain, Abnormal nuclear pore formation triggers apoptosis in the intestinal epithelium of elys-deficient zebrafish. 2009, Pubmed
Devos, Components of coated vesicles and nuclear pore complexes share a common molecular architecture. 2004, Pubmed
Doucet, Cell cycle-dependent differences in nuclear pore complex assembly in metazoa. 2010, Pubmed , Xenbase
Drummond, Interference with the cytoplasmic tail of gp210 disrupts "close apposition" of nuclear membranes and blocks nuclear pore dilation. 2002, Pubmed , Xenbase
Dultz, Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells. 2008, Pubmed
Dultz, Live imaging of single nuclear pores reveals unique assembly kinetics and mechanism in interphase. 2010, Pubmed
Earp, The many mechanisms of viral membrane fusion proteins. 2005, Pubmed
Eriksson, Dynamic properties of nuclear pore complex proteins in gp210 deficient cells. 2004, Pubmed
Feldherr, The location of the transport gate in the nuclear pore complex. 1997, Pubmed , Xenbase
Fernandez, MEL-28 is downstream of the Ran cycle and is required for nuclear-envelope function and chromatin maintenance. 2006, Pubmed
Fort, Comparative sensitivity of Xenopus tropicalis and Xenopus laevis as test species for the FETAX model. 2004, Pubmed , Xenbase
Franz, MEL-28/ELYS is required for the recruitment of nucleoporins to chromatin and postmitotic nuclear pore complex assembly. 2007, Pubmed , Xenbase
Fridkin, Matefin, a Caenorhabditis elegans germ line-specific SUN-domain nuclear membrane protein, is essential for early embryonic and germ cell development. 2004, Pubmed
Funakoshi, Two distinct human POM121 genes: requirement for the formation of nuclear pore complexes. 2007, Pubmed
Galy, MEL-28, a novel nuclear-envelope and kinetochore protein essential for zygotic nuclear-envelope assembly in C. elegans. 2006, Pubmed
Gattegno, Genetic control of fusion pore expansion in the epidermis of Caenorhabditis elegans. 2007, Pubmed
Gerace, Identification of a major polypeptide of the nuclear pore complex. 1982, Pubmed
Gillespie, ELYS/MEL-28 chromatin association coordinates nuclear pore complex assembly and replication licensing. 2007, Pubmed , Xenbase
Goldberg, Dimples, pores, star-rings, and thin rings on growing nuclear envelopes: evidence for structural intermediates in nuclear pore complex assembly. 1997, Pubmed , Xenbase
Goldstein, Pleiotropic nuclear defects associated with a conditional allele of the novel nucleoporin Rat9p/Nup85p. 1996, Pubmed
Greber, A major glycoprotein of the nuclear pore complex is a membrane-spanning polypeptide with a large lumenal domain and a small cytoplasmic tail. 1990, Pubmed
Griffis, Nup98 localizes to both nuclear and cytoplasmic sides of the nuclear pore and binds to two distinct nucleoporin subcomplexes. 2003, Pubmed
Gruenbaum, The nuclear lamina comes of age. 2005, Pubmed
Güttinger, Orchestrating nuclear envelope disassembly and reassembly during mitosis. 2009, Pubmed
Hallberg, An integral membrane protein of the pore membrane domain of the nuclear envelope contains a nucleoporin-like region. 1993, Pubmed
Harel, Removal of a single pore subcomplex results in vertebrate nuclei devoid of nuclear pores. 2003, Pubmed , Xenbase
Harel, Importin beta negatively regulates nuclear membrane fusion and nuclear pore complex assembly. 2003, Pubmed , Xenbase
Henderson, The temperature arrested intermediate of virus-cell fusion is a functional step in HIV infection. 2006, Pubmed
Herrmann, Protein-mediated phospholipid translocation in the endoplasmic reticulum with a low lipid specificity. 1990, Pubmed
Hetzer, GTP hydrolysis by Ran is required for nuclear envelope assembly. 2000, Pubmed , Xenbase
Hetzer, Border control at the nucleus: biogenesis and organization of the nuclear membrane and pore complexes. 2009, Pubmed
Higa, Studying nuclear disassembly in vitro using Xenopus egg extract. 2006, Pubmed , Xenbase
Hinshaw, Architecture and design of the nuclear pore complex. 1992, Pubmed , Xenbase
Hodge, Integral membrane proteins Brr6 and Apq12 link assembly of the nuclear pore complex to lipid homeostasis in the endoplasmic reticulum. 2010, Pubmed
Holaska, Multiple roles for emerin: implications for Emery-Dreifuss muscular dystrophy. 2006, Pubmed
Hu, Membrane proteins of the endoplasmic reticulum induce high-curvature tubules. 2008, Pubmed
Huebner, Nurse cell-oocyte interaction: a new F-actin mesh associated with the microtubule-rich core of an insect ovariole. 1986, Pubmed
Jackson, Fusion pores and fusion machines in Ca2+-triggered exocytosis. 2006, Pubmed
Jahn, Membrane fusion. 2003, Pubmed
Kalab, The RanGTP gradient - a GPS for the mitotic spindle. 2008, Pubmed
Kalab, Visualization of a Ran-GTP gradient in interphase and mitotic Xenopus egg extracts. 2002, Pubmed , Xenbase
Katsani, In vivo dynamics of Drosophila nuclear envelope components. 2008, Pubmed
Keminer, Permeability of single nuclear pores. 1999, Pubmed , Xenbase
Kielian, Virus membrane-fusion proteins: more than one way to make a hairpin. 2006, Pubmed
Kozlov, [Possible mechanism of membrane fusion]. 1983, Pubmed
Kutay, Reorganization of the nuclear envelope during open mitosis. 2008, Pubmed
Lang, Molecular mobility and nucleocytoplasmic flux in hepatoma cells. 1986, Pubmed
Lau, Topology of yeast Ndc1p: predictions for the human NDC1/NET3 homologue. 2006, Pubmed
Lau, Transportin regulates major mitotic assembly events: from spindle to nuclear pore assembly. 2009, Pubmed , Xenbase
Lénárt, Nuclear envelope breakdown in starfish oocytes proceeds by partial NPC disassembly followed by a rapidly spreading fenestration of nuclear membranes. 2003, Pubmed
Li, Mutation or deletion of the Saccharomyces cerevisiae RAT3/NUP133 gene causes temperature-dependent nuclear accumulation of poly(A)+ RNA and constitutive clustering of nuclear pore complexes. 1995, Pubmed
Liu, The COPI complex functions in nuclear envelope breakdown and is recruited by the nucleoporin Nup153. 2003, Pubmed , Xenbase
Liu, Functional association of Sun1 with nuclear pore complexes. 2007, Pubmed
Lohka, Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components. 1983, Pubmed , Xenbase
Lu, Membrane fusion induced by neuronal SNAREs transits through hemifusion. 2005, Pubmed
Lutzmann, Modular self-assembly of a Y-shaped multiprotein complex from seven nucleoporins. 2002, Pubmed
Macaulay, Assembly of the nuclear pore: biochemically distinct steps revealed with NEM, GTP gamma S, and BAPTA. 1996, Pubmed , Xenbase
Madrid, The role of the integral membrane nucleoporins Ndc1p and Pom152p in nuclear pore complex assembly and function. 2006, Pubmed
Mansfeld, The conserved transmembrane nucleoporin NDC1 is required for nuclear pore complex assembly in vertebrate cells. 2006, Pubmed , Xenbase
Markosyan, HIV-1 envelope proteins complete their folding into six-helix bundles immediately after fusion pore formation. 2003, Pubmed
Marx, Rapid flip-flop of phospholipids in endoplasmic reticulum membranes studied by a stopped-flow approach. 2000, Pubmed
Mayer, Membrane fusion in eukaryotic cells. 2002, Pubmed
Melia, Lipidic antagonists to SNARE-mediated fusion. 2006, Pubmed
Mische, Direct observation of regulated ribonucleoprotein transport across the nurse cell/oocyte boundary. 2007, Pubmed
Mishra, The Nup107-160 complex and gamma-TuRC regulate microtubule polymerization at kinetochores. 2010, Pubmed , Xenbase
Mohler, The type I membrane protein EFF-1 is essential for developmental cell fusion. 2002, Pubmed
Naim, Passive and facilitated transport in nuclear pore complexes is largely uncoupled. 2007, Pubmed
Newport, The nucleus: structure, function, and dynamics. 1987, Pubmed
Newport, Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. 1987, Pubmed , Xenbase
Newport, Characterization of the membrane binding and fusion events during nuclear envelope assembly using purified components. 1992, Pubmed , Xenbase
Olsson, Limited expression of nuclear pore membrane glycoprotein 210 in cell lines and tissues suggests cell-type specific nuclear pores in metazoans. 2004, Pubmed
Onischenko, Role of the Ndc1 interaction network in yeast nuclear pore complex assembly and maintenance. 2009, Pubmed
Orjalo, The Nup107-160 nucleoporin complex is required for correct bipolar spindle assembly. 2006, Pubmed , Xenbase
Papadopulos, Flippase activity detected with unlabeled lipids by shape changes of giant unilamellar vesicles. 2007, Pubmed
Peters, Nuclear envelope permeability measured by fluorescence microphotolysis of single liver cell nuclei. 1983, Pubmed
Peters, Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis. 1984, Pubmed
Podbilewicz, The C. elegans developmental fusogen EFF-1 mediates homotypic fusion in heterologous cells and in vivo. 2006, Pubmed
Rafikova, Transmembrane protein-free membranes fuse into xenopus nuclear envelope and promote assembly of functional pores. 2009, Pubmed , Xenbase
Rasala, ELYS is a dual nucleoporin/kinetochore protein required for nuclear pore assembly and proper cell division. 2006, Pubmed , Xenbase
Rasala, Capture of AT-rich chromatin by ELYS recruits POM121 and NDC1 to initiate nuclear pore assembly. 2008, Pubmed , Xenbase
Ratner, Molecular determinants of binding between Gly-Leu-Phe-Gly nucleoporins and the nuclear pore complex. 2007, Pubmed
Reese, Trans-SNARE pairing can precede a hemifusion intermediate in intracellular membrane fusion. 2005, Pubmed
Rotem, Importin beta regulates the seeding of chromatin with initiation sites for nuclear pore assembly. 2009, Pubmed , Xenbase
Sapir, AFF-1, a FOS-1-regulated fusogen, mediates fusion of the anchor cell in C. elegans. 2007, Pubmed
Saraste, Temperature-sensitive steps in the transport of secretory proteins through the Golgi complex in exocrine pancreatic cells. 1986, Pubmed
Scarcelli, The yeast integral membrane protein Apq12 potentially links membrane dynamics to assembly of nuclear pore complexes. 2007, Pubmed
Schneiter, A yeast acetyl coenzyme A carboxylase mutant links very-long-chain fatty acid synthesis to the structure and function of the nuclear membrane-pore complex. 1996, Pubmed
Schoch, A long-lived state for influenza virus-erythrocyte complexes committed to fusion at neutral pH. 1992, Pubmed
Schwander, Beta1 integrins regulate myoblast fusion and sarcomere assembly. 2003, Pubmed
Shibata, The reticulon and DP1/Yop1p proteins form immobile oligomers in the tubular endoplasmic reticulum. 2008, Pubmed , Xenbase
Siniossoglou, Structure and assembly of the Nup84p complex. 2000, Pubmed
Sondermann, Prediction of novel Bag-1 homologs based on structure/function analysis identifies Snl1p as an Hsp70 co-chaperone in Saccharomyces cerevisiae. 2002, Pubmed
Stavru, Nuclear pore complex assembly and maintenance in POM121- and gp210-deficient cells. 2006, Pubmed
Stavru, NDC1: a crucial membrane-integral nucleoporin of metazoan nuclear pore complexes. 2006, Pubmed
Stegmann, Intermediates in influenza induced membrane fusion. 1990, Pubmed
Stewart, Blurring the boundary: the nuclear envelope extends its reach. 2007, Pubmed
Taimen, A progeria mutation reveals functions for lamin A in nuclear assembly, architecture, and chromosome organization. 2009, Pubmed
Tzur, SUN-domain proteins: 'Velcro' that links the nucleoskeleton to the cytoskeleton. 2006, Pubmed
Vasu, Novel vertebrate nucleoporins Nup133 and Nup160 play a role in mRNA export. 2001, Pubmed , Xenbase
Voeltz, A class of membrane proteins shaping the tubular endoplasmic reticulum. 2006, Pubmed
Walther, The conserved Nup107-160 complex is critical for nuclear pore complex assembly. 2003, Pubmed , Xenbase
Walther, RanGTP mediates nuclear pore complex assembly. 2003, Pubmed , Xenbase
Wozniak, Primary structure analysis of an integral membrane glycoprotein of the nuclear pore. 1989, Pubmed
Xu, Hemifusion in SNARE-mediated membrane fusion. 2005, Pubmed
Zhang, Chromatin-independent nuclear envelope assembly induced by Ran GTPase in Xenopus egg extracts. 2000, Pubmed , Xenbase