Functional Translatome Proteomics Reveal Converging and Dose-Dependent Regulation by mTORC1 and eIF2α

Mol Cell. 2020 Feb 20;77(4):913-925.e4. doi: 10.1016/j.molcel.2019.11.010. Epub 2019 Dec 4.


Regulation of translation is essential during stress. However, the precise sets of proteins regulated by the key translational stress responses-the integrated stress response (ISR) and mTORC1-remain elusive. We developed multiplexed enhanced protein dynamics (mePROD) proteomics, adding signal amplification to dynamic-SILAC and multiplexing, to enable measuring acute changes in protein synthesis. Treating cells with ISR/mTORC1-modulating stressors, we showed extensive translatome modulation with ∼20% of proteins synthesized at highly reduced rates. Comparing translation-deficient sub-proteomes revealed an extensive overlap demonstrating that target specificity is achieved on protein level and not by pathway activation. Titrating cap-dependent translation inhibition confirmed that synthesis of individual proteins is controlled by intrinsic properties responding to global translation attenuation. This study reports a highly sensitive method to measure relative translation at the nascent chain level and provides insight into how the ISR and mTORC1, two key cellular pathways, regulate the translatome to guide cellular survival upon stress.

Keywords: SILAC; TMT; cap-dependent translation; integrated stress response; mTOR; proteomics; pulse labeling; stress response; translation; unfolded protein response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Eukaryotic Initiation Factor-2 / metabolism*
  • HeLa Cells
  • Humans
  • Mechanistic Target of Rapamycin Complex 1 / metabolism*
  • Protein Biosynthesis*
  • Proteomics / methods*
  • Stress, Physiological / genetics
  • Unfolded Protein Response


  • Eukaryotic Initiation Factor-2
  • Mechanistic Target of Rapamycin Complex 1