TY - JOUR
T1 - Functional Proteomics Identifies Acinus L as a Direct Insulin- and Amino Acid-Dependent Mammalian Target of Rapamycin Complex 1 (mTORC1) Substrate
AU - Schwarz, Jennifer Jasmin
AU - Wiese, Heike
AU - Toelle, Regine Charlotte
AU - Zarei, Mostafa
AU - Dengjel, Joern
AU - Warscheid, Bettina
AU - Thedieck, Kathrin
N1 - © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2015/8
Y1 - 2015/8
N2 - The serine/threonine kinase mammalian target of rapamycin (mTOR) governs growth, metabolism, and aging in response to insulin and amino acids (aa), and is often activated in metabolic disorders and cancer. Much is known about the regulatory signaling network that encompasses mTOR, but surprisingly few direct mTOR substrates have been established to date. To tackle this gap in our knowledge, we took advantage of a combined quantitative phosphoproteomic and interactomic strategy. We analyzed the insulin- and aa-responsive phosphoproteome upon inhibition of the mTOR complex 1 (mTORC1) component raptor, and investigated in parallel the interactome of endogenous mTOR. By overlaying these two datasets, we identified acinus L as a potential novel mTORC1 target. We confirmed acinus L as a direct mTORC1 substrate by co-immunoprecipitation and MS-enhanced kinase assays. Our study delineates a triple proteomics strategy of combined phosphoproteomics, interactomics, and MS-enhanced kinase assays for the de novo-identification of mTOR network components, and provides a rich source of potential novel mTOR interactors and targets for future investigation.
AB - The serine/threonine kinase mammalian target of rapamycin (mTOR) governs growth, metabolism, and aging in response to insulin and amino acids (aa), and is often activated in metabolic disorders and cancer. Much is known about the regulatory signaling network that encompasses mTOR, but surprisingly few direct mTOR substrates have been established to date. To tackle this gap in our knowledge, we took advantage of a combined quantitative phosphoproteomic and interactomic strategy. We analyzed the insulin- and aa-responsive phosphoproteome upon inhibition of the mTOR complex 1 (mTORC1) component raptor, and investigated in parallel the interactome of endogenous mTOR. By overlaying these two datasets, we identified acinus L as a potential novel mTORC1 target. We confirmed acinus L as a direct mTORC1 substrate by co-immunoprecipitation and MS-enhanced kinase assays. Our study delineates a triple proteomics strategy of combined phosphoproteomics, interactomics, and MS-enhanced kinase assays for the de novo-identification of mTOR network components, and provides a rich source of potential novel mTOR interactors and targets for future investigation.
KW - NOVO PYRIMIDINE SYNTHESIS
KW - EXON JUNCTION COMPLEX
KW - RICH AKT SUBSTRATE
KW - P70 S6 KINASE
KW - 40 KDA PRAS40
KW - CELL-GROWTH
KW - CHROMATIN CONDENSATION
KW - MOTIF PHOSPHORYLATION
KW - SIGNAL INTEGRATION
KW - BINDING PARTNER
U2 - 10.1074/mcp.M114.045807
DO - 10.1074/mcp.M114.045807
M3 - Article
C2 - 25907765
VL - 14
SP - 2042
EP - 2055
JO - Molecular & Cellular Proteomics
JF - Molecular & Cellular Proteomics
SN - 1535-9476
IS - 8
ER -