FIGURE 2.

Pcl1 is more stable in the absence of Dma activity. A, Pcl1 levels increased in the presence of proteasomal inhibitors. Cells of strain YSH82 were incubated with either MG132 (a proteasome inhibitor) or a drug vehicle (DMSO). 60 min later, samples were taken and Pcl1-TAP protein levels were analyzed by immunoblotting using monoclonal antibodies against TAP-tag. Two different exposures of the immunoblot are shown. As in the rest of the panels, Glu-6-PDH (G6PDH) detection was used as a loading control. B, relative amounts of Pcl1 in wild type, grr1Δ, and dma1Δ dma2Δ strains. Cells were grown exponentially in rich media, and Pcl1-TAP levels were detected by immunoblotting using monoclonal antibodies. C, Pcl1 stability measurements. The indicated strains were grown exponentially in rich media and cycloheximide was added to the medium at time 0. Samples were taken at the indicated times and analyzed for Pcl1-TAP levels by immunoblotting using monoclonal antibodies. D, quantification of panel C. Data ± S.D. from 3 independent experiments are shown. *, p > 0.05 versus WT. E, Pcl9 stability measurements were carried out as in panel C. F, quantification of panel E. Data ± S.D. from 3 independent experiments are shown.