Yeast Culture Compound Screening

The following molecules were incubated in yeast cultures for 10 min at the following concentrations. Rapamycin (TORC1 inhibitor, 200 ng/mL - Fisher Scientific BP2963-1), dissolved in DMSO (Amresco 0231); 1,10 Phenanthroline (RNAP I–III inhibitor, 100 µg/mL - Sigma-Aldrich 131377), in ethanol (Fisher Scientific BP28184); doxycycline (Antibiotic/regulator of Tet-Off/On promoters; 10 µg/mL - Sigma-Aldrich D-9891) in water, β-estradiol (regulator of β-estradiol promoter; 1 µM - Sigma-Aldrich E8875), in ethanol; thiolutin (RNAP I-III inhibitor, 3 µg/mL - VWR 89157-260), in DMSO; 4tU (Uracil analog, 25 µg/mL - Sigma-Aldrich 440736), in DMSO. All drug concentrations are based on standards in the field and/or the following studies (Grigull et al. 2004 (link); Mnaimneh et al. 2004 (link); Munchel et al. 2011 (link)). Ten minutes was chosen as a time period for analysis given that mRNA half-lives were recently reported to average 5 min (Chan et al. 2018 (link)), thus short-term effects are relevant for accurate half-life measurement.