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Altering nuclear pore complex function impacts longevity and mitochondrial function in S. cerevisiae.

Lord CL, Timney BL, Rout MP, Wente SR - J. Cell Biol. (2015)

Bottom Line: Mutants lacking the GLFG domain of Nup116 displayed decreased RLSs, whereas longevity was increased in nup100- mutants.Both Kap121-dependent transport and Nup116 levels decrease in replicatively aged yeast.Together, these studies reveal that specific NPC nuclear transport events directly influence aging.

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Affiliation: Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232.

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Specific GLFG domains regulate RLS independently of NPC permeability. (A and B) Survival curves of wild-type and mutant cells (n ≥ 50) grown at 30°C. Mean RLSs are listed in parentheses next to genotypes. *, P < 0.001 when the curve is compared with wild type using a log-rank test. (C) Mean half-times for GFP-4PrA equilibration after cytoplasmic bleaching in the listed strains. *, P < 0.01 using Tukey’s post-hoc test when compared with wild-type cells after a one-way analysis of variance (ANOVA; n ≥ 10 cells per strain). Error bars represent SEM. (D) Representative time-lapse FRAP images of single cells expressing GFP-4PrA at the listed times relative to the bleach. Bar, 2.5 µm. (E) A list of mutant strains and their mean RLSs. *, P < 0.05 when the survival curve is compared with wild type using a log-rank test with n ≥ 50 cells.
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fig1: Specific GLFG domains regulate RLS independently of NPC permeability. (A and B) Survival curves of wild-type and mutant cells (n ≥ 50) grown at 30°C. Mean RLSs are listed in parentheses next to genotypes. *, P < 0.001 when the curve is compared with wild type using a log-rank test. (C) Mean half-times for GFP-4PrA equilibration after cytoplasmic bleaching in the listed strains. *, P < 0.01 using Tukey’s post-hoc test when compared with wild-type cells after a one-way analysis of variance (ANOVA; n ≥ 10 cells per strain). Error bars represent SEM. (D) Representative time-lapse FRAP images of single cells expressing GFP-4PrA at the listed times relative to the bleach. Bar, 2.5 µm. (E) A list of mutant strains and their mean RLSs. *, P < 0.05 when the survival curve is compared with wild type using a log-rank test with n ≥ 50 cells.

Mentions: Based on previous studies showing the composition and functions of NPCs change as cells and organisms age (D’Angelo et al., 2009; Kelley et al., 2011), we hypothesized altering NPC function might affect RLS in S. cerevisiae. To test this, we focused on measuring the RLSs of a subset of NPC mutant strains lacking specific FG domains (ΔFG; Strawn et al., 2004; Terry and Wente, 2007). The mean RLS of nup116ΔGLFG mutants was decreased ∼20% when compared with wild-type W303 cells (Fig. 1 A). Although RLS was unaffected in nup145ΔGLFG cells, mean RLS was further reduced 20% in nup116ΔGLFG nup145ΔGLFG double mutants (Fig. 1 A). Surprisingly, deletion of NUP100 (nup100Δ) or its GLFG domain (nup100ΔGLFG) significantly increased mean RLS 17% in W303 (Fig. 1 B) and 23% in BY4741 cells (Fig. S1 A). In contrast, RLS was not significantly altered in nsp1ΔFG, nup145ΔGLFG nup100ΔGLFG, or ΔN ΔC (a strain lacking the five FG domains from Nup42, Nup159, Nup1, Nup2, and Nup60) mutants (Fig. 1 E). Immunoblotting of total cell lysates showed the levels of Nup116 and Nup100 were not significantly altered in nup116ΔGLFG nup145ΔGLFG and nup100ΔGLFG cells (Fig. S1, C and D). Together, these results suggested there are specific defects associated with nup116ΔGLFG, nup116ΔGLFG nup145ΔGLFG, and nup100ΔGLFG cells that alter RLS, and demonstrated longevity is regulated by NPC structure and function in S. cerevisiae.


Altering nuclear pore complex function impacts longevity and mitochondrial function in S. cerevisiae.

Lord CL, Timney BL, Rout MP, Wente SR - J. Cell Biol. (2015)

Specific GLFG domains regulate RLS independently of NPC permeability. (A and B) Survival curves of wild-type and mutant cells (n ≥ 50) grown at 30°C. Mean RLSs are listed in parentheses next to genotypes. *, P < 0.001 when the curve is compared with wild type using a log-rank test. (C) Mean half-times for GFP-4PrA equilibration after cytoplasmic bleaching in the listed strains. *, P < 0.01 using Tukey’s post-hoc test when compared with wild-type cells after a one-way analysis of variance (ANOVA; n ≥ 10 cells per strain). Error bars represent SEM. (D) Representative time-lapse FRAP images of single cells expressing GFP-4PrA at the listed times relative to the bleach. Bar, 2.5 µm. (E) A list of mutant strains and their mean RLSs. *, P < 0.05 when the survival curve is compared with wild type using a log-rank test with n ≥ 50 cells.
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Related In: Results  -  Collection

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fig1: Specific GLFG domains regulate RLS independently of NPC permeability. (A and B) Survival curves of wild-type and mutant cells (n ≥ 50) grown at 30°C. Mean RLSs are listed in parentheses next to genotypes. *, P < 0.001 when the curve is compared with wild type using a log-rank test. (C) Mean half-times for GFP-4PrA equilibration after cytoplasmic bleaching in the listed strains. *, P < 0.01 using Tukey’s post-hoc test when compared with wild-type cells after a one-way analysis of variance (ANOVA; n ≥ 10 cells per strain). Error bars represent SEM. (D) Representative time-lapse FRAP images of single cells expressing GFP-4PrA at the listed times relative to the bleach. Bar, 2.5 µm. (E) A list of mutant strains and their mean RLSs. *, P < 0.05 when the survival curve is compared with wild type using a log-rank test with n ≥ 50 cells.
Mentions: Based on previous studies showing the composition and functions of NPCs change as cells and organisms age (D’Angelo et al., 2009; Kelley et al., 2011), we hypothesized altering NPC function might affect RLS in S. cerevisiae. To test this, we focused on measuring the RLSs of a subset of NPC mutant strains lacking specific FG domains (ΔFG; Strawn et al., 2004; Terry and Wente, 2007). The mean RLS of nup116ΔGLFG mutants was decreased ∼20% when compared with wild-type W303 cells (Fig. 1 A). Although RLS was unaffected in nup145ΔGLFG cells, mean RLS was further reduced 20% in nup116ΔGLFG nup145ΔGLFG double mutants (Fig. 1 A). Surprisingly, deletion of NUP100 (nup100Δ) or its GLFG domain (nup100ΔGLFG) significantly increased mean RLS 17% in W303 (Fig. 1 B) and 23% in BY4741 cells (Fig. S1 A). In contrast, RLS was not significantly altered in nsp1ΔFG, nup145ΔGLFG nup100ΔGLFG, or ΔN ΔC (a strain lacking the five FG domains from Nup42, Nup159, Nup1, Nup2, and Nup60) mutants (Fig. 1 E). Immunoblotting of total cell lysates showed the levels of Nup116 and Nup100 were not significantly altered in nup116ΔGLFG nup145ΔGLFG and nup100ΔGLFG cells (Fig. S1, C and D). Together, these results suggested there are specific defects associated with nup116ΔGLFG, nup116ΔGLFG nup145ΔGLFG, and nup100ΔGLFG cells that alter RLS, and demonstrated longevity is regulated by NPC structure and function in S. cerevisiae.

Bottom Line: Mutants lacking the GLFG domain of Nup116 displayed decreased RLSs, whereas longevity was increased in nup100- mutants.Both Kap121-dependent transport and Nup116 levels decrease in replicatively aged yeast.Together, these studies reveal that specific NPC nuclear transport events directly influence aging.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232.

Show MeSH
Related in: MedlinePlus