Skip to main content
. 2021 Jan 30;296:100207. doi: 10.1074/jbc.RA120.014956

Figure 7.

Figure 7

eIF2B localization in the presence of gcn3-containing vanishing whie matter (VWM) missense mutations.A, polysome analysis of the strain yMK1402 (GCD1-yeGFPgcn3::LEU2) expressing gcn3 VWM mutants and the low-copy WTGCN3 plasmid (ySC91–96). Polysome analysis was as described in Experimental procedures. Polysome/monosome ratios were calculated from measuring the area under the polysome peaks and dividing by the monosome peak area. B, (i) cells were grown to log phase, and confocal microscopy was used to image the strain yMK1402 (GCD1-yeGFPgcn3::LEU2) expressing gcn3 VWM mutants, Gcn3pK11E (p[GCN3K111EURA3CEN6ARS4]), Gcn3pV184D (p[GCN3V184DURA3CEN6ARS4]), Gcn3pN209Y (p[GCN3N209YURA3CEN6ARS4]), Gcn3pF230V (p[GCN3F240VURA3CEN6ARS4]), Gcn3pY274C (p[GCN3Y274CURA3CEN6ARS4]), and the low-copy WT plasmid p[GCN3URA3CEN6ARS4]. (ii) cells from each strain were counted to assess whether eIF2B bodies were present, dispersed, or localized to microfoci. For each strain analyzed, the localization of eIF2B was assessed for 100 cells per replicate, n = 3. Error bars are representative of SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. ns, not significant; SCD, synthetic complete media.