FIGURE 6:

Suppressees of Sec61-2-GFP lethality are ChrV and XIV aneuploids. (A) Constitutive overexpression of Sec61-2-GFP is lethal to asi1Δhrd1Δ cells. Left, schematic denoting the genotypes of each strain tested before exposure to 5-FOA. Center and right, the indicated strains were streaked onto plates that either selected (-Trp -Ura) or counterselected the URA3 plasmids. Plates were incubated at 30°C for 2 d before imaging. One representative of three biological replicates is shown. (B) Lethality suppressees cannot degrade the Sec61-2-GFP. Four suppressees and a WT strain expressing Sec61-2-GFP were subjected to CHX chase. After the addition of CHX, cells were assayed for fluorescence by flow cytometry, and at each time point, the mean fluorescence of 10,000 cells was measured. t = 0 was taken as 100%, and data plotted are the mean ± SD from three experiments. (C) Genome profiling reveals duplications of ChrV and XIV in suppressees. Chromosome profiles of whole-genome sequencing are mapped across the yeast genome. Copy number is indicated on the y-axis, and the chromosome number is indicated on the x-axis. Reads from each of four suppressees are shown.