FIG. 4.

The superkiller rpl3 mutants do not confer defects on the ability of mutant ribosomes to recruit the Ski-complex to mRNAs. JD1090 cells expressing pRPL3 (WT) or the L17S, P18S, or Y283C alleles were transformed with pAJ1203 (FLAG-tagged SKI7). WT-c represents wild-type cells transformed with a vector control. Cycloheximide-arrested ribosomes were purified by ultracentrifugation from logarithmically growing cells, separated through sodium dodecyl sulfate-PAGE, transferred to PVDF membranes, and probed using anti-FLAG or anti-L3 antiserum. Locations of the Ski7-FLAG and L3 are indicated. Anti-FLAG-reactive nonspecific bands are indicated by an asterisk.