Genomic Run On (GRO): determination of the nascent transcriptional rates in a series of Hrp1 mutants.. Saccharomyces cerevisiae

The goal of the project was to check the differential effect on transcription rate of a sudden degron-induced depletion of Hpr1 protein and its comparison towards the effect of a constitutive absence of Hpr1 protein in a deletion strain. Overall design: Cells were grown in YP-Raffinose + 0.1 mM CuSO4 at 26ºC to 0.5 OD600. At that cells were spun-down and changed to YP-Galactose without CuSO4 for 30 min at 26ºC. Then cells were shifted to 37ºC to induce degron-mediated proteolysis and recovered after 30 min. Samples include wt, hpr1delta and hpr1-degron after 0 or 30 min growth at 30ºC.