FIG. 5.

Induced transcription. (A) Derepression of SUC2 and ADH2. Cultures of YJR241-x were grown initially in YPAD at 23°C. Following the removal of an aliquot from each culture, cells were centrifuged, resuspended in YPA containing a low concentration of dextrose (0.05%) prewarmed to 37°C, and grown at the restrictive temperature. RNA was isolated from aliquots that were removed after 1.5 and 3 h at 37°C in low-dextrose medium. Transcripts were detected by Northern blotting. scR1 was used as a loading control. (B) Analysis of a DNA damage-induced gene. Cultures of YJR241-x were grown in rich medium and shifted to the nonpermissive temperature. Aliquots were removed (permissive [lanes P]) prior to the temperature shift. After 1 h at 37°C, MMS was added to 0.02%. Aliquots were taken at 1.5 and 3 h after treatment with MMS. A separate culture was maintained at 37°C but left untreated (nonpermissive [lanes N]) for 4 h. HUG1 RNA was analyzed by Northern blotting. scR1 was used as a loading control.