During development, morphogens provide extracellular cues allowing cells to select a specific fate by inducing complex transcriptional programs. The mating pathway in budding yeast offers simplified settings to understand this process. Pheromone secreted by the mating partner triggers the activity of a MAPK pathway, which results in the expression of hundreds of genes. Using a dynamic expression reporter, we quantified the kinetics of gene expression in single cells upon exogenous pheromone stimulation and in the physiological context of mating. In both conditions, we observed striking differences in the timing of induction of mating‐responsive promoters. Biochemical analyses and generation of synthetic promoter variants demonstrated how the interplay between transcription factor binding and nucleosomes contributes to determine the kinetics of transcription in a simplified cell‐fate decision system. Synopsis Quantitative and dynamic single cell measurements uncover a complex temporal orchestration of gene expression events in the yeast mating response. The promoter architecture controls the timing of gene expression relative to the time of fusion. Exogenous stimulations of yeast cells with pheromone have allowed to classify mating promoters in three categories, based on their gene expression dynamics. The number and affinity of transcription factor binding sites in nucleosome depleted regions regulate the timing of gene induction. In presence of mating partners, early genes are expressed during the sensing phase, while late genes are induced shortly before fusion. Quantitative and dynamic single cell measurements uncover a complex temporal orchestration of gene expression events in the yeast mating response. The promoter architecture controls the timing of gene expression relative to the time of fusion.