A comprehensive temporal proteome and phosphoproteome dataset of Y1 adrenocortical cells under FGF2 stimulation and SOS inhibition

Abstract

The Y1 mouse adrenocortical tumor cell line is a classic model for cancer studies and ACTH mediated steroidogenesis. The KRAS gene amplification and SOS mediated activation sustain elevated ERK levels, which drive the tumor phenotype of this cell line. In addition, the mitogen FGF2 induces atypical cytotoxic stress and consequent cell death rather than proliferation in this system, imposing a strong perturbation on KRAS ERK signaling dynamics. Recent research from our group revealed that SOS is not solely responsible for KRAS activation in the Y1 cells, indicating a more complex regulatory landscape of GEFs and GAPs. This motivated the generation of a comprehensive time resolved proteome (DIA) and TMT phosphoproteome (DDA) in a 0 to 30 minute time course under FGF2 stimulation or combined with SOS inhibitor, with the presence or absence of FBS stimulation, in technical triplicate using Q Exactive and Thermo Lumos mass spectrometers, respectively. This dataset can enable latent mechanisms between the life and death switch caused by FGF2 signaling and the alternate stress response caused by impaired SOS dependent activation. This analysis yielded 14,512 phosphosites and 8,919 protein groups, where 10,471 phosphosites and 111 protein groups showed differential abundance with statistical significance (limma, FDR < 0.05%). This dataset provides a versatile resource for downstream applications, which includes dynamic modeling such as ordinary differential equation based frameworks, for which temporal datasets remain scarce. The data contained in this preprint will be available on JPOST Repo after the publication.