Yeast cell growth and -1/+1 PRF monitoring

Dilution spot assays were used to qualitatively monitor cell growth at 30, 37 and 15°C. For all conditions, yeast cells were grown to logarithmic growth phase and then diluted to 10⁶ CFU/ml. Subsequently, 10-fold serial dilutions of each strain were spotted onto YPAD. The dual luciferase reporter plasmids pYDL-control, pYDL-LA, pYDL-Ty1, pYDL-UAA (34 (link)), pYDL-AGC₂₁₈ and pYDL-TCT₂₁₈ (35 (link)) were employed to monitor L-A-directed programmed -1 ribosomal frameshifting (-1 PRF), Ty1-directed +1 PRF, UAA termination codon readthrough and suppression of an AGC near-cognate serine codon or a TCT noncognate serine codon in the firefly luciferase catalytic site, respectively. All assays were performed in quadruplicate and were repeated a minimum of four times or until the statistical requirements were met (36 (link)). Sample readings were collected using a GloMax Multi-Microplate luminometer (Promega).