Figure 2.
RPN11 is required for the UV- and MMS-induced expression of Gcn4p target genes. (A) UV-induced expression of HIS3 and HIS4. Wild-type and mutant cells (JR493) were treated on plates with 70 J/m2 UV and returned to prewarmed YPAD for 75 min. HIS3 and ACT1 mRNAs were detected by S1 nuclease protection, and HIS4 mRNA by primer extension. +1 and +13 indicate the two major transcription start sites of HIS3. (B) MMS-induced induction of HIS3, HIS4, and HIS5. Cells were treated with (+) or without (−) 0.1% MMS for 1 h at 30°C. HIS5 mRNA was detected by Northern blotting. (C) Examination of HIS3 and HIS4 expression under amino acid starvation. Cells were grown at 30°C in liquid SC media (−) or in media lacking histidine and treated with 15 mM 3-AT for 60 min (+). RNA was analyzed as described in A.