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Example sentences for: nup
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Logarithmically growing cultures of npa1-1/sec23 and npa2-1/sec13 were shifted to the non-permissive temperature as before and then fixed and processed for IF with polyclonal antibodies against Kar2p to visualize the ER [ 45 ] . The monoclonal antibody mAb414, which recognizes a subset of Nups distinct from Nic96p and Nup170p [ 46 ] , was also used because the GFP fluorescence of the GFP-Nups was more sensitive to fixation when the GFP-Nups were mislocalized than when they remained at the nuclear rim (unpublished observation).
Wild type GFP-nic96 nup170-GFP haploid strains were mutagenized to generate a bank of ts mutant strains (Figure 1A; see Materials and Methods).
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.
Nup62 was detectable in smaller amounts in the cytosol and NP fractions as well (Fig 3B), consistent with its cellular distribution described previously [ 30 ] . On the other hand, Hsp110 was found predominantly in the cytosol fraction, while Hsp90 was found both in the cytosol and the NP fractions (Fig 3B).
Like GFP-Nic96p and Nup170p-GFP, the localization of Nup116p and Pom152p changed from a punctate, nuclear rim staining to cytoplasmic foci when shifted to the non-permissive temperature (data not shown).
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