Temporal Proteomic Profiling of Pheromone-Induced Cell Cycle Re-Entry in Saccharomyces cerevisiae

The regulation of cell cycle progression in response to environmental cues is essential for cellular adaptation. In Saccharomyces cerevisiae, the BAR1 gene modulates sensitivity to the mating pheromone α-factor, which induces cell cycle arrest. Here we investigated the dynamic proteomic response in the bar1 deletion strain using a 27-plex experimental design with TMTproD isobaric labeling. Asynchronous bar1Δ cells were arrested with α-factor and then released from the pheromone arrest. We acquired three replicate protein abundance time-course profiles following pheromone (α-factor) washout, with samples collected at eight time points from 0 to 165 minutes post-washout. Using higher-order TMTpro sample multiplexing, we generated global temporal profiles of protein abundance associated with recovery from pheromone-induced arrest. Our findings identify specific proteins and pathways involved in cell cycle re-entry and in the attenuation of the pheromone signal, providing insights into the regulatory mechanisms of mating response in yeast. This study contributes significantly to dynamic proteomic analysis in cell cycle progression. We present a powerful approach for investigating complex cellular processes and showcase cell cycle progression following pheromone washout in yeast.