Δard1 yeast have a growth defect, but no translocation defect or Sec62p instability. (A) Wildtype (BY4742) and the indicated mutant cells were diluted on YPD plates and growth monitored at the indicated temperatures. (B) Wildtype, sec61-302 and Δard1 cells were transformed with empty vector (pRS313), or plasmids expressing PRC1-URA3 (CPY) or PHO8-URA3 (PHO8) fusions. Growth in absence (−URA) or presence (+URA) of uracil was monitored on plates at 30°C. Growth in the absence of uracil indicates a defect in translocation into the ER. (C) Wildtype (BY4742) and the indicated mutant cells were grown to early log phase, cycloheximide added at time 0, and samples taken at the indicated chase times. Cells were lysed, proteins resolved by SDS PAGE and Sec62p and Pdi1p (as loading control) detected by immunoblotting.