FIG. 4.
Transcriptional analysis of the K151L,K156Y strain under restrictive conditions indicates defects for a collection of Pol II-transcribed genes. (A) Mutant and wild-type (wt) TBP-containing strains were grown to log phase and shifted to the restrictive temperature for 1 h. Total RNA isolated before and after the temperature shift was hybridized with a 100-fold excess of the indicated probe and treated with S1 nuclease; tRNAW served as a loading control. The promoter for the +1 transcript from the HIS3 gene is considered TATA-less, while the promoter for the +13 transcript from the HIS3 gene contains a canonical TATA element. The promoters of the other genes tested (MOT1, CMD1, RPS4, ADH1, PGK1, DED1, ENO2, HTA2, and FUR1) have recognizable TATA elements within 250 bp of their translation start sites. The functional relevance of many of these elements is not currently known. (B) Analysis of cyclin genes at the restrictive temperature. Total RNAs from the indicated strains grown at 30°C and for 1 h at 38°C were blotted to nylon membranes and subsequently hybridized with the indicated cyclin probes. ADH1 served as a loading and transfer control.