FIG. 5.

(A) Sirtuin regulation of TNA1, TNR1, and TNR2. S1 nuclease analysis of TNA1, TNR1/TNR2, and ACT1 transcript levels. RNA was prepared from five strains, the wild type (WT; BG2) and the hst1 (BG1218), hst2 (BG1219), sir2 (BG1216), sir4 (BG1217) mutants. RNA was prepared from cells grown in SC medium containing 3.25 μM NA to an OD₆₀₀ of 0.5. (B) S1 nuclease analysis of TNA1, TNR1/TNR2, and ACT1 transcript levels. RNA was prepared from three strains, the wild type (BG2) and the hst1 (BG1218) and sum1 (BG1462) mutants, growing in SC medium containing 3.25 μM (lanes 1, 3, and 5) or 16 nM (lanes 2, 4, and 6) NA. The OD₆₀₀ of all of the cultures was 0.5. (C) S1 nuclease analysis of EPA6, TNR1/TNR2, and ACT1 transcript levels. Strain BG2 was grown in SC medium containing the indicated initial concentrations of NA; all cultures were grown to an OD₆₀₀ of 0.5.