Figure 7.

Rad51-3A is degraded by the 26S proteasome. Exponential cultures of indicated strains were treated with MMS (0.02%) 30 min prior to the addition of cycloheximide (200 μg/ml) and/or MG132 (25 μM). Protein samples were harvested at indicated time-points and immunoblotting was performed as described in Figure 6. To decrease MG132 efflux, we deleted the PDR5 gene, which encodes an ABC transporter. The protein levels of another vegetative house-keeping protein, hexokinase, were analyzed by immunoblotting using anti-hexokinase antibodies and are shown as the loading control in (A). Quantification of immunoblotting results in (A) and (D) are plotted in (C) and (E), respectively, in which total Rad51 protein levels normalized to Hsp104 levels at that time-point are plotted. Quantification results of samples of WT and rad51-3A with cycloheximide from (C) were included in (E) for comparison.