Example sentences for: npcs
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This strategy was based on the ability to functionally express GFP-nucleoporin fusions, and the assumption that NPC a ssembly/mislocalization mutants expressing a GFP-nucleoporin would have distinct fluorescence properties [ 26 ] . In wild type yeast cells, fluorescence microscopy localization of proteins at NPCs appears as punctate staining of virtually the entire nuclear rim circumference (see Figure 2).
Both regions contained multiple open reading frames; however, we focused immediately on a single gene for each mutant, SEC13 for npa2-1 and SEC23 for npa1-1 . SEC13 was first identified in a screen for secretory mutants and encodes a protein of the COPII coat, which is required for vesicle budding from the ER [ 37 38 39 ] . More recently, it has been shown that a fraction of the total cellular Sec13p co-fractionates with NPCs in a distinct subcomplex with Nup84p, Nup85p, Nup120p, Nup145Cp and Seh1p [ 40 41 ] . SEC23 was also identified in a screen for secretory mutants, and it, too, is a component of the COPII coat [ 37 38 39 ] . Co-purification of Sec23p with intact NPCs has not been reported [ 5 ] ; however, Sec23p was recently isolated in Nup42p and Nup116p affinity chromatography strategies [ 42 ] . Expression constructs containing only SEC13 or SEC23 were transformed into the mutant strains and assayed for their ability to rescue growth at the non-permissive temperature.
After staining with osmium tetroxide and uranyl acetate, NPCs in wild-type/parental cells were represented by electron dense structures spanning the pore (Figure 4).
Using GFP-tagged NPCs, we designed live cell assays to monitor NPC dynamics and assembly rates [ 25 ] . Moreover, to isolate factors required for proper NPC assembly and localization, we developed a novel genetic screening strategy.
De novo assembly into intact NEs occurs during cell and nuclear growth to maintain a constant NPC density [ 10 ] . In cells undergoing an open mitosis, NPCs are also re-assembled with the formation of the NE at the end of mitosis [ 11 ] . The mechanisms controlling membrane fusion during pore formation and the steps for assembly of the distinct NPC substructures have not been fully elucidated.
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