Skip to main content
. 2010 May 1;24(9):893–903. doi: 10.1101/gad.1906510

Figure 1.

Figure 1.

SLX5 and SLX8 are required for degradation of the α2*-Ura3-3HA reporter protein. (A) Schematic of the two major ubiquitin conjugation pathways that regulate α2 degradation. (B) Growth assay for α2*-Ura3-3HA metabolic stabilization. The slx5Δ and slx8Δ mutants were transformed with a centromeric plasmid carrying the α2*-URA3-3HA allele, grown in liquid medium lacking leucine (to maintain the LEU2 plasmid), and then spotted on both SD-leucine and SD-uracil plates in 10-fold serial dilutions. The α2* protein carries the I4T and L10S substitutions, which inhibit degradation by the Ubc6/7–Doa10 pathway. Pictures were taken after 1 d (SD-leucine) or 3 d (SD-uracil) at 30°C. (C) Cycloheximide-chase/immunoblot analysis of α2*-Ura3-3HA degradation. Cell extracts from the indicated strains were harvested at the indicated times after addition of cycloheximide, and were resolved by 8% SDS-PAGE followed by anti-HA immunoblotting. Anti-Pgk1immunoblotting allowed comparison of protein loading between samples. (α2*-UH) α2*-Ura3-3HA.