Figure 1

(A) Yeast plasmids pRep41/(nmt promoter medium expression, LEU+) and pRep42 (nmt promoter medium expression, Ura+) containing human glucocorticoid receptor sequences (GRα) and the various deletant mutants. (B) Western blot analysis of yeast strains transformed with empty vector (WT), or human GR full sequences or disruption mutants. Yeast were grown in EMM selective medium to mid-long phase. Protein lysates were probed for GRα antibody. (C) Thermotolerance of transformed yeast. S. pombe were transformed with the empty plasmid pRep42 (WT), or with full length human GRα or the disruption mutants as indicated. The transformants were grown in selective EMM. Serially diluted yeast (5 fold dilution series) was spotted on selective minimal media and incubated for 5 days at 30 °C or 39  °C. Results are representative of three independent experiments. (D) Summary diagram showing the required N (AF-1) and C (Ligand binding domain, LBD) terminal domains to confer thermoprotection in yeast.