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. 2012 Jan 1;23(1):137–150. doi: 10.1091/mbc.E11-05-0419

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© 2012 Garre et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — The cbc1Δ mutant shows a defect in translation recovery after hyperosmotic stress. (A) Reinitiation of translation after adaptation to osmotic stress is delayed in the cbc1Δ mutant. Polysome profiles show translation inhibition and recovery during osmotic stress in the wt, hog1Δ, and cbc1Δ strains. Cells grown exponentially in YPD were treated with 0.6 M KCl at the indicated times. Polysomal profiles were obtained as described in Materials and Methods. The relative amount of rRNA in each fraction, measured by online UV detection at 260 nm, shows the ribosomal light subunits (40S), heavy subunits (60S), monosomes (80S), and polysomes (2n, 3n, 4n, …). Representative absorbance curves (out of five repetitions) are displayed. (B) Percentage of the polysomal area during osmotic stress (0.6 M KCl) in wt, hog1Δ, and cbc1Δ. The polysome percentage is relative to the polysomal area of each strain at time 0. The data are given as the mean and SE of three independent experiments. (C) P-body assembly and disassembly under osmotic shock are delayed in the cbc1Δ mutant. The wt, hog1Δ, and cbc1Δ strains expressing a GFP-tagged version of the decapping enzyme Dcp2 were grown until the exponential phase, and then treated with 1 M KCl, 0.6 M KCl, or with 0.4 M NaCl. Dcp2-GFP allowed P-body visualization by fluorescence microscopy. (D) The quantification of the percentage of cells with P-bodies is indicated in the graphs. At least 100 cells were quantified for each condition.