Proximity-dependent biotin labeling (BioID) may identify new targets for cancers driven by difficult-to-drug oncogenes such as Ras. Therefore， BioID was used with wild-type (WT) and oncogenic mutant (MT) H-，?K-， and N-Ras， identifying known interactors， including Raf and PI3K， as well as a common set of 130 novel proteins proximal to all Ras isoforms. A CRISPR screen of these proteins for Ras dependence identified mTOR， which was also found proximal to MT Ras in human tumors. Oncogenic Ras directly bound two mTOR complex 2 (mTORC2) components， mTOR and MAPKAP1， to promote mTORC2 kinase activity at the plasma membrane. mTORC2 enabled the Ras pro-proliferative cell cycle?transcriptional program... More
Proximity-dependent biotin labeling (BioID) may identify new targets for cancers driven by difficult-to-drug oncogenes such as Ras. Therefore， BioID was used with wild-type (WT) and oncogenic mutant (MT) H-，?K-， and N-Ras， identifying known interactors， including Raf and PI3K， as well as a common set of 130 novel proteins proximal to all Ras isoforms. A CRISPR screen of these proteins for Ras dependence identified mTOR， which was also found proximal to MT Ras in human tumors. Oncogenic Ras directly bound two mTOR complex 2 (mTORC2) components， mTOR and MAPKAP1， to promote mTORC2 kinase activity at the plasma membrane. mTORC2 enabled the Ras pro-proliferative cell cycle?transcriptional program， and perturbing the Ras-mTORC2 interaction impaired Ras-dependent neoplasia in?vivo. Combining proximity-dependent proteomics with CRISPR screening identified a new set of functional Ras-associated proteins， defined mTORC2 as a new direct Ras effector， and offers a strategy for finding new proteins that cooperate with dominant oncogenes.
