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. 2002 Aug;22(15):5395–5404. doi: 10.1128/MCB.22.15.5395-5404.2002

FIG. 8.

FIG. 8.

Pcl5 disappears rapidly in starved cells and is constitutively unstable. (A) Western blot with 50 μg of extract from pcl5Δ cells or from W303-1A cells overexpressing Pcl5 from plasmid KB1093 and either grown in SC medium (LOG), shifted for 30 min to YNB medium plus adenine (STARV), or incubated for 30 min with cycloheximide at 5 μg/ml (CHX). The arrow indicates Pcl5; the star indicates across-reacting band. (B) Pulse-chase analysis of Pcl5 overexpressed from the KB1093 plasmid either in normal (SC) medium or after a shift for 30 min to YNB plus adenine (SD) medium. Note that the same amount of total incorporated radioactivity was processed for each time point; therefore, the signal strength at time zero is not indicative of total cellular protein biosynthesis. (C) Degradation of endogenous Pcl5. W303-1A cells were subjected to pulse-chase analysis after induction of PCL5 expression by Gcn4 expressed from the KB843 plasmid. pcl5Δ strain KY827 was used as a control. The arrow indicates the position of the Pcl5 band. kD, kilodaltons.