Fig. 7

Collided ribosomes are associated with specific chaperones. a The schematic of detecting the disome-enriched proteins through SILAC. Two biological replicates were performed with reciprocal labeling (the left panel). Because equal amount of total protein extracted from the monosome fraction and disome fraction were mixed, proteins having the same abundance per ribosome between monosomes and disomes should be located on the diagonal (the right panel). Proteins locating above or below the diagonal should be those enriched in the disomes or the monosomes, respectively. b The protein intensities per ribosome in disomes were plotted against those in monosomes. Chaperones (green) are enriched in the disome, whereas most constitutive ribosomal proteins (pink) show the same abundance per ribosome between monosomes and disomes. c The m/z (mass divided by charge number) spectrum of Ssb1p and its fragments detected in MS/MS. d The enrichment of chaperones in disomes. Immunoblotting was performed using anti-FLAG antibody, with Ponceau S stained total protein as the loading control. Mono and Di represent proteins extracted from the monosome and disome fractions, respectively. e A schematic of the fates of ribosome collisions with various durations. The double-rotated disome conformation is used to show the initial status for ribosome collisions (left). The signal peptide is highlighted in orange. SRP, signal recognition particle; ER, endoplasmic reticulum; Ub, ubiquitin