Mammalian cells respond to amino acid limitation by activating a coordinated response to preserve cell function and resources. Here we dissected amino acid stress sensing and adaptation and examined how prolonged arginine or leucine starvation alters cellular behavior of murine embryonic stem cells, that served as a model rapidly proliferating, non-transformed system. We used a temporal approach that combined proteomics, phosphoproteomics (EasyPhos), ubiqitinomics (TUBE-MS), and interaction proteomics (AP-MS) to provide a systematic investigation of the nutrient stress response and regulation networks of mTORC1 (with Castor1 and Sesn2 sensors) as well as cellular adaptation mechanisms.