Figure 2.

Identification of two sets of stress genes differentially regulated by Sty1-Atf1. A, stress-dependent transcriptional analysis of Atf1 phosphomutants. YE cultures of strains 972 (WT), EP201 (Δatf1 + emp.), EP203 (Δatf1 + HA-Atf1), EP203.10D (Δatf1 + HA-Atf1.10D), and EP203.10M (Δatf1 + HA-Atf1.10M) were treated with 1 mm H₂O₂ for 15 min or left untreated. Total RNA was analyzed by Northern blotting as described in Fig. 1D. B, expression of phospho-mimicking HA-Atf1.10D suppresses the transcription defects of cells lacking Sty1. Cultures of strains 972 (WT), EP288 (Δatf1 Δsty1 + emp.), EP303 (Δatf1 Δsty1 + HA-Atf1), EP303.10D (Δatf1 Δsty1 + HA-Atf1.10D) and EP303.10M (Δatf1 Δsty1 + HA-Atf1.10M) were treated with 1 mm H₂O₂ for 15 min, and total RNA was analyzed as described in Fig. 1D. C, expression of HA-Atf1.10D suppresses the sensitivity to peroxides of cells lacking Sty1. The same cultures as in B were spotted on plates containing H₂O₂ as described in Fig. 1B. D, representation of 246 genes whose transcript levels increase by at least 2-fold in wild-type cells upon treatment with 1 mm H₂O₂ for 15 min. Total RNA from strains 972 (WT), AV18 (Δsty1), EP303, EP303.10D, and EP303.10M (Δatf1 Δsty1 + HA-Atf1, HA-Atf1.10D, and HA-Atf1.10M, respectively) was sequenced using Illumina technology and analyzed as described under “Experimental Procedures.” Horizontal strips represent genes, and columns represent untreated or treated conditions of the indicated strains. The log2 changes in expression, relative to the untreated wild-type sample, are color-coded as shown in the bar. Genes were hierarchically clustered based on the expression patterns of cells expressing HA-Atf1.10D. The relative positions of some representative genes (srx1, ctt1, gpd1, and hsp9) are indicated on the right.