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. 2019 Aug 1;33(15-16):1031–1047. doi: 10.1101/gad.327494.119

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© 2019 Brennan et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 3. — Stoichiometric imbalances of protein complex subunits cause protein aggregation. (A) The percentage of proteins encoded by duplicated chromosomes enriched by log₂ ≥1.27 in aggregates (red), the percentage of proteins encoded by euploid chromosomes enriched by log₂ ≥1.27 in aggregates (gray), and the percentage of proteins in the genome (black) that are annotated to form protein complexes by Pu et al. (2009) were calculated. (****) P < 0.0001; (n.s.) not significant, cumulative distribution function for a binomial distribution. (B) Diagram of eIF2 complex stoichiometry in disome V cells. (C) Gcd11-HA in aggregates and total lysates in WT (A40189), disome V (A40190), disome V GCD11-HA/gcd11Δ (A40191), and WT URA3::GCD11-HA (WT + GCD11; A40192) cells. Only one of the two GCD11 genes in disome V cells was tagged with HA to ensure that protein levels are comparable between strains. (D) Quantification of Western blots in C. n = 3; SD. (***) P < 0.001, t-test. (E) Cells were grown as in C, and mRNA levels for eIF2 subunits were determined. Values were normalized to ACT1 and then to WT expression levels. n = 3; SD. (F–H) Western blot, relative aggregation quantification, and mRNA expression for WT (A40193), disome V (A40194), and disome V pSUI2/SUI3 (A40195) cells as described in C–E. n = 3; SD. (**) P < 0.01, t-test. (Dis) Disome; (rel) relative.