Genetic interactions of BRL1. (A) Indicated yeast strains with chromosomal PADH1-BBR6 or PADH1-BRL1 or plasmid encoded PGAL1-BRR6 or PGAL1-BRL1 were incubated for 2 d at 37°C. –, no growth; +, growth; –/+, partial growth; nd, not determined. A cartoon of the NPC with indicated proteins tested in the assay is shown. (B) Serial dilutions of indicated strains with the plasmid-encoded BRL1 alleles on SC-selection plates. (C) Suppression of the growth defect of nup116ΔGLFG PMET3-NUP188 cells by PADH1-BRL1. Ten-fold serial dilutions of cells on plates with or without methionine. (D) Growth of brr6(ts), brl1(ts), and nup116Δ cells with PADH1-NUP116. (E) TEM images of nup116Δ and gle2Δ cells show herniation defects after incubation at 37°C for 3 and 2.5 h, respectively. N, nucleus; C, cytoplasm. Bars: (overviews) 500 nm; (enlargements) 100 nm. (F) Immuno-EM analysis with anti-GFP antibody. nup116Δ and gle2Δ cells with PADH1-yeGFP-BRL1 were incubated for 3 and 2.5 h at 37°C, respectively. Arrowheads, 10-nm gold particles reflecting yeGFP localization. Abbreviations and bars are as in E.
