Figure 6. Defective ribosome assembly homeostasis and proteostatic collapse in tom1 mutant cells.

(A–C) Hypersensitivity of tom1^CA cells to imbalances in ribosome components. (A) Cells of the indicated genotypes were spotted in serial 10-fold dilutions on glucose or galactose medium and incubated at 30°C for 2 days. ev refers to empty vector. n = 2 biological replicates. (B, C) As in (A) except that cells of the indicated genotypes were spotted on YPD. n = 2 biological replicates. (D–G) Massive accumulation of insoluble proteins in tom1 mutant cells. Cells of the indicated genotypes were lysed and fractionated into detergent-soluble and insoluble fractions, which were separated by SDS-PAGE and stained with Coomassie Blue. The pellet fraction is overloaded 20-fold compared to the total and supernatant fractions. n = 2 biological replicates.

DOI: http://dx.doi.org/10.7554/eLife.19105.015

Figure 6—figure supplement 1. Tom1 is required for maintaining proteostasis.

(A) Cells of the indicated genotypes were spotted in serial 10-fold dilutions on glucose or galactose medium and incubated at 30°C for 2 days. ev refers to empty vector. n = 2 biological replicates. (B) Cells of the indicated genotypes were spotted as serial 10-fold dilutions on YPD with or without 1 mM auxin and incubated at 30°C for 2 days. AID refers to auxin-inducible degron and CA refers to the Cys3235Ala mutation in TOM1. n = 2 biological replicates. (C) List of genetic interactions with tom1∆ as reported in the Saccharomyces Genome Database (www.yeastgenome.org). (D) Synthetic growth defects of rps29a∆tom1∆ double mutants. Cells of the indicated genotypes were spotted in serial 10-fold dilutions on YPD and incubated at 30°C for 2 days. n = 2 biological replicates. (E) same as panel B except medium was supplemented or not with 2 mM paromomycin.

DOI: http://dx.doi.org/10.7554/eLife.19105.016