Updated project metadata. We systematically exposed the yeast Saccharomyces cerevisiae to a wide array of 101 perturbations and generated a quantitative mass spectrometry-based atlas of early phosphoproteomic responses. Perturbations covered broad classes of environmental, chemical, and drug perturbations and systematically targeted diverse aspects of yeast cell biology. Importantly, most perturbations focused on capturing early signaling responses (5 min treatments), with the reasoning that functional phosphorylation of specific substrates occurs rapidly following stimulation, but promiscuous phosphorylation and confounding changes in transcript and protein abundances takes place more slowly. Treatments were done in 6 biological replicates and large-scale phosphoproteomics was enabled by our recently developed R2-P2 (Rapid-Robotic Phosphoproteomics) sample preparation workflow in combination with DIA measurements. This submission includes 559 quantitative phospho DIA-MS raw files that went into the analysis of the systems-level phosphorylation stress response.