Figure 3.—

Additive effects of Δtor1 and Δtsc2 mutations. (A) Wild-type (TA001), Δtor1 (TA390), Δtsc2 (TA450), and Δtor1 Δtsc2 (TA459) cells were grown to midlog phase in EMM (M) medium, washed, resuspended in proline (P) medium, and further incubated for 15 min at 30°. RNA was extracted and subjected to Northern blot (top two sections) or semiquantitative RT–PCR analyses (bottom two sections). Northern blots were probed with isp5⁺ and adh1⁺ (loading control). For RT–PCR analysis, primers were specific for the amino acid permease 7G5.06 or FKBP12 (control). (B) Δtor1 (TA390), Δtsc2 (TA450), and Δtor1 Δtsc2 (TA459) cells were grown to midlog phase in EMM medium. Incorporation of l-³H-leucine was measured after 6 min using the protocol detailed in materials and methods.