{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T16:19:08Z","timestamp":1774628348715,"version":"3.50.1"},"reference-count":39,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2016,6,14]],"date-time":"2016-06-14T00:00:00Z","timestamp":1465862400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2016,6,14]],"date-time":"2016-06-14T00:00:00Z","timestamp":1465862400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>It is well known that the GCN2 and mTORC1 signaling pathways are regulated by amino acids and share common functions, in particular the control of translation. The regulation of GCN2 activity by amino acid availability relies on the capacity of GCN2 to sense the increased levels of uncharged tRNAs upon amino acid scarcity. In contrast, despite recent progress in the understanding of the regulation of mTORC1 by amino acids, key aspects of this process remain unsolved. In particular, while leucine is well known to be a potent regulator of mTORC1, the mechanisms by which this amino acid is sensed and control mTORC1 activity are not well defined. Our data establish that GCN2 is involved in the inhibition of mTORC1 upon leucine or arginine deprivation. However, the activation of GCN2 alone is not sufficient to inhibit mTORC1 activity, indicating that leucine and arginine exert regulation via additional mechanisms. While the mechanism by which GCN2 contributes to the initial step of mTORC1 inhibition involves the phosphorylation of eIF2\u03b1, we show that it is independent of the downstream transcription factor ATF4. These data point to a novel role for GCN2 and phosphorylation of eIF2\u03b1 in the control of mTORC1 by certain amino acids.<\/jats:p>","DOI":"10.1038\/srep27698","type":"journal-article","created":{"date-parts":[[2016,6,14]],"date-time":"2016-06-14T09:53:12Z","timestamp":1465897992000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":85,"title":["GCN2 contributes to mTORC1 inhibition by leucine deprivation through an ATF4 independent mechanism"],"prefix":"10.1038","volume":"6","author":[{"given":"Julien","family":"Averous","sequence":"first","affiliation":[]},{"given":"Sarah","family":"Lambert-Langlais","sequence":"additional","affiliation":[]},{"given":"Florent","family":"Mesclon","sequence":"additional","affiliation":[]},{"given":"Val\u00e9rie","family":"Carraro","sequence":"additional","affiliation":[]},{"given":"Laurent","family":"Parry","sequence":"additional","affiliation":[]},{"given":"C\u00e9line","family":"Jousse","sequence":"additional","affiliation":[]},{"given":"Alain","family":"Bruhat","sequence":"additional","affiliation":[]},{"given":"Anne-Catherine","family":"Maurin","sequence":"additional","affiliation":[]},{"given":"Philippe","family":"Pierre","sequence":"additional","affiliation":[]},{"given":"Christopher G.","family":"Proud","sequence":"additional","affiliation":[]},{"given":"Pierre","family":"Fafournoux","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2016,6,14]]},"reference":[{"key":"BFsrep27698_CR1","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/S0092-8674(02)00808-5","volume":"110","author":"D-H Kim","year":"2002","unstructured":"Kim, D.-H. et al. mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery. Cell 110, 163\u2013175 (2002).","journal-title":"Cell"},{"key":"BFsrep27698_CR2","doi-asserted-by":"publisher","first-page":"1432","DOI":"10.1073\/pnas.95.4.1432","volume":"95","author":"PE Burnett","year":"1998","unstructured":"Burnett, P. E., Barrow, R. K., Cohen, N. A., Snyder, S. H. & Sabatini, D. M. RAFT1 phosphorylation of the translational regulators p70 S6 kinase and 4E-BP1. Proc. Natl. Acad. Sci. USA 95, 1432\u20131437 (1998).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"BFsrep27698_CR3","doi-asserted-by":"publisher","first-page":"14484","DOI":"10.1074\/jbc.273.23.14484","volume":"273","author":"K Hara","year":"1998","unstructured":"Hara, K. et al. Amino Acid Sufficiency and mTOR Regulate p70 S6 Kinase and eIF-4E BP1 through a Common Effector Mechanism. J. Biol. Chem. 273, 14484\u201314494 (1998).","journal-title":"J. Biol. Chem."},{"key":"BFsrep27698_CR4","doi-asserted-by":"publisher","first-page":"754","DOI":"10.1046\/j.1432-1327.1999.00780.x","volume":"265","author":"JJ Berlanga","year":"1999","unstructured":"Berlanga, J. J., Santoyo, J. & de Haro, C. Characterization of a mammalian homolog of the GCN2 eukaryotic initiation factor 2alpha kinase. Eur. J. Biochem. 265, 754\u2013762 (1999).","journal-title":"Eur. J. Biochem."},{"key":"BFsrep27698_CR5","doi-asserted-by":"publisher","first-page":"11269","DOI":"10.1073\/pnas.0400541101","volume":"101","author":"KM Vattem","year":"2004","unstructured":"Vattem, K. M. & Wek, R. C. Reinitiation involving upstream ORFs regulates ATF4 mRNA translation in mammalian cells. Proc Natl Acad Sci USA 101, 11269\u201374 (2004).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep27698_CR6","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1042\/BJ20060941","volume":"402","author":"AB Lopez","year":"2007","unstructured":"Lopez, A. B. et al. A feedback transcriptional mechanism controls the level of the arginine\/lysine transporter cat-1 during amino acid starvation. Biochem. J. 402, 163\u2013173 (2007).","journal-title":"Biochem. J."},{"key":"BFsrep27698_CR7","doi-asserted-by":"publisher","first-page":"24120","DOI":"10.1074\/jbc.M201959200","volume":"277","author":"F Siu","year":"2002","unstructured":"Siu, F., Bain, P. J., LeBlanc-Chaffin, R., Chen, H. & Kilberg, M. S. ATF4 is a mediator of the nutrient-sensing response pathway that activates the human asparagine synthetase gene. J Biol Chem 277, 24120\u20137 (2002).","journal-title":"J Biol Chem"},{"key":"BFsrep27698_CR8","doi-asserted-by":"publisher","first-page":"7683","DOI":"10.1093\/nar\/gkt563","volume":"41","author":"W B\u2019Chir","year":"2013","unstructured":"B\u2019Chir, W. et al. The eIF2alpha\/ATF4 pathway is essential for stress-induced autophagy gene expression. Nucleic Acids Res 41, 7683\u201399 (2013).","journal-title":"Nucleic Acids Res"},{"key":"BFsrep27698_CR9","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1097\/MCO.0000000000000240","volume":"19","author":"DJ Ham","year":"2016","unstructured":"Ham, D. J., Lynch, G. S. & Koopman, R. Amino acid sensing and activation of mechanistic target of rapamycin complex 1: implications for skeletal muscle. Curr. Opin. Clin. Nutr. Metab. Care 19, 67\u201373 (2016).","journal-title":"Curr. Opin. Clin. Nutr. Metab. Care"},{"key":"BFsrep27698_CR10","doi-asserted-by":"publisher","first-page":"1496","DOI":"10.1126\/science.1157535","volume":"320","author":"Y Sancak","year":"2008","unstructured":"Sancak, Y. et al. The Rag GTPases Bind Raptor and Mediate Amino Acid Signaling to mTORC1. Science 320, 1496\u20131501 (2008).","journal-title":"Science"},{"key":"BFsrep27698_CR11","doi-asserted-by":"publisher","first-page":"290","DOI":"10.1016\/j.cell.2010.02.024","volume":"141","author":"Y Sancak","year":"2010","unstructured":"Sancak, Y. et al. Ragulator-Rag Complex Targets mTORC1 to the Lysosomal Surface and is Necessary for its Activation by Amino Acids. Cell 141, 290\u2013303 (2010).","journal-title":"Cell"},{"key":"BFsrep27698_CR12","doi-asserted-by":"publisher","first-page":"1918","DOI":"10.1016\/j.cellsig.2014.04.019","volume":"26","author":"J Averous","year":"2014","unstructured":"Averous, J. et al. Requirement for lysosomal localization of mTOR for its activation differs between leucine and other amino acids. Cell. Signal. 26, 1918\u20131927 (2014).","journal-title":"Cell. Signal."},{"key":"BFsrep27698_CR13","doi-asserted-by":"publisher","first-page":"754","DOI":"10.1016\/j.ccell.2014.09.008","volume":"26","author":"JD Thomas","year":"2014","unstructured":"Thomas, J. D. et al. Rab1A is an mTORC1 Activator and a Colorectal Oncogene. Cancer Cell 26, 754\u2013769 (2014).","journal-title":"Cancer Cell"},{"key":"BFsrep27698_CR14","doi-asserted-by":"publisher","first-page":"C206","DOI":"10.1152\/ajpcell.1998.274.1.C206","volume":"274","author":"HL Fox","year":"1998","unstructured":"Fox, H. L., Kimball, S. R., Jefferson, L. S. & Lynch, C. J. Amino acids stimulate phosphorylation of p70S6k and organization of rat adipocytes into multicellular clusters. Am J Physiol 274, C206\u201313 (1998).","journal-title":"Am J Physiol"},{"key":"BFsrep27698_CR15","doi-asserted-by":"publisher","first-page":"269","DOI":"10.1016\/S1097-2765(00)00028-9","volume":"6","author":"J Dong","year":"2000","unstructured":"Dong, J., Qiu, H., Garcia-Barrio, M., Anderson, J. & Hinnebusch, A. G. Uncharged tRNA activates GCN2 by displacing the protein kinase moiety from a bipartite tRNA-binding domain. Mol Cell 6, 269\u201379 (2000).","journal-title":"Mol Cell"},{"key":"BFsrep27698_CR16","doi-asserted-by":"publisher","first-page":"746","DOI":"10.2337\/db10-1246","volume":"60","author":"F Xiao","year":"2011","unstructured":"Xiao, F. et al. Leucine Deprivation Increases Hepatic Insulin Sensitivity via GCN2\/mTOR\/S6K1 and AMPK Pathways. Diabetes 60, 746\u2013756 (2011).","journal-title":"Diabetes"},{"key":"BFsrep27698_CR17","doi-asserted-by":"publisher","first-page":"36553","DOI":"10.1074\/jbc.M404559200","volume":"279","author":"TG Anthony","year":"2004","unstructured":"Anthony, T. G. et al. Preservation of Liver Protein Synthesis during Dietary Leucine Deprivation Occurs at the Expense of Skeletal Muscle Mass in Mice Deleted for eIF2 Kinase GCN2. J. Biol. Chem. 279, 36553\u201336561 (2004).","journal-title":"J. Biol. Chem."},{"key":"BFsrep27698_CR18","doi-asserted-by":"publisher","first-page":"2709","DOI":"10.1016\/j.cellsig.2013.08.038","volume":"25","author":"MD Dennis","year":"2013","unstructured":"Dennis, M. D., McGhee, N. K., Jefferson, L. S. & Kimball, S. R. Regulated in DNA damage and development 1 (REDD1) promotes cell survival during serum deprivation by sustaining repression of signaling through the mechanistic target of rapamycin in complex 1 (mTORC1). Cell. Signal 25, 2709\u20132716 (2013).","journal-title":"Cell. Signal"},{"key":"BFsrep27698_CR19","doi-asserted-by":"publisher","first-page":"11106","DOI":"10.1128\/JVI.01063-07","volume":"81","author":"K Minami","year":"2007","unstructured":"Minami, K. et al. Suppression of viral replication by stress-inducible GADD34 protein via the mammalian serine\/threonine protein kinase mTOR pathway. J. Virol. 81, 11106\u201311115 (2007).","journal-title":"J. Virol."},{"key":"BFsrep27698_CR20","doi-asserted-by":"publisher","first-page":"29900","DOI":"10.1074\/jbc.M003633200","volume":"275","author":"S Mordier","year":"2000","unstructured":"Mordier, S., Deval, C., Bechet, D., Tassa, A. & Ferrara, M. Leucine limitation induces autophagy and activation of lysosome-dependent proteolysis in C2C12 myotubes through a mammalian target of rapamycin-independent signaling pathway. J Biol Chem 275, 29900\u20136 (2000).","journal-title":"J Biol Chem"},{"key":"BFsrep27698_CR21","doi-asserted-by":"publisher","first-page":"1099","DOI":"10.1016\/S1097-2765(00)00108-8","volume":"6","author":"HP Harding","year":"2000","unstructured":"Harding, H. P. et al. Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell 6, 1099\u2013108. (2000).","journal-title":"Mol Cell"},{"key":"BFsrep27698_CR22","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1042\/bj20021266","volume":"372","author":"A Beugnet","year":"2003","unstructured":"Beugnet, A., Tee, A. R., Taylor, P. M. & Proud, C. G. Regulation of targets of mTOR (mammalian target of rapamycin) signalling by intracellular amino acid availability. Biochem. J. 372, 555\u2013566 (2003).","journal-title":"Biochem. J."},{"key":"BFsrep27698_CR23","doi-asserted-by":"crossref","first-page":"3854","DOI":"10.1016\/S0021-9258(19)45112-0","volume":"247","author":"BS Hansen","year":"1972","unstructured":"Hansen, B. S., Vaughan, M. H. & Wang, L. Reversible inhibition by histidinol of protein synthesis in human cells at the activation of histidine. J. Biol. Chem. 247, 3854\u20133857 (1972).","journal-title":"J. Biol. Chem."},{"key":"BFsrep27698_CR24","doi-asserted-by":"publisher","first-page":"6515","DOI":"10.1128\/MCB.00489-09","volume":"29","author":"C Chaveroux","year":"2009","unstructured":"Chaveroux, C. et al. Identification of a novel amino acid response pathway triggering ATF2 phosphorylation in mammals. Mol Cell Biol 29, 6515\u201326 (2009).","journal-title":"Mol Cell Biol"},{"key":"BFsrep27698_CR25","doi-asserted-by":"publisher","first-page":"4843","DOI":"10.1073\/pnas.0735876100","volume":"100","author":"J Lee","year":"2003","unstructured":"Lee, J., Ryu, H., Ferrante, R. J., Morris, S. M. & Ratan, R. R. Translational control of inducible nitric oxide synthase expression by arginine can explain the arginine paradox. Proc. Natl. Acad. Sci. 100, 4843\u20134848 (2003).","journal-title":"Proc. Natl. Acad. Sci"},{"key":"BFsrep27698_CR26","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1038\/cr.2015.146","volume":"26","author":"M Shimobayashi","year":"2016","unstructured":"Shimobayashi, M. & Hall, M. N. Multiple amino acid sensing inputs to mTORC1. Cell Res. 26, 7\u201320 (2016).","journal-title":"Cell Res."},{"key":"BFsrep27698_CR27","doi-asserted-by":"publisher","first-page":"31827","DOI":"10.1074\/jbc.M114.602870","volume":"289","author":"M Sokabe","year":"2014","unstructured":"Sokabe, M. & Fraser, C. S. Human Eukaryotic Initiation Factor 2 (eIF2)-GTP-Met-tRNAi Ternary Complex and eIF3 Stabilize the 43 S Preinitiation Complex. J. Biol. Chem. 289, 31827\u201331836 (2014).","journal-title":"J. Biol. Chem."},{"key":"BFsrep27698_CR28","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1016\/j.cell.2005.10.024","volume":"123","author":"MK Holz","year":"2005","unstructured":"Holz, M. K., Ballif, B. A., Gygi, S. P. & Blenis, J. mTOR and S6K1 mediate assembly of the translation preinitiation complex through dynamic protein interchange and ordered phosphorylation events. Cell 123, 569\u2013580 (2005).","journal-title":"Cell"},{"key":"BFsrep27698_CR29","doi-asserted-by":"crossref","unstructured":"Tsokanos, F.-F. et al. eIF4A inactivates TORC1 in response to amino acid starvation. EMBO J. n\/a\u2013n\/a. doi: 10.15252\/embj.201593118 (2016).","DOI":"10.15252\/embj.201593118"},{"key":"BFsrep27698_CR30","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1126\/science.1257132","volume":"347","author":"S Wang","year":"2015","unstructured":"Wang, S. et al. Lysosomal amino acid transporter SLC38A9 signals arginine sufficiency to mTORC1. Science 347, 188\u2013194 (2015).","journal-title":"Science"},{"key":"BFsrep27698_CR31","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1038\/nature14107","volume":"519","author":"M Rebsamen","year":"2015","unstructured":"Rebsamen, M. et al. SLC38A9 is a component of the lysosomal amino acid-sensing machinery that controls mTORC1. Nature 519, 477\u2013481 (2015).","journal-title":"Nature"},{"key":"BFsrep27698_CR32","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1016\/j.cell.2012.02.044","volume":"149","author":"JM Han","year":"2012","unstructured":"Han, J. M. et al. Leucyl-tRNA Synthetase is an Intracellular Leucine Sensor for the mTORC1-Signaling Pathway. Cell 149, 410\u2013424 (2012).","journal-title":"Cell"},{"key":"BFsrep27698_CR33","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1016\/j.molcel.2012.05.043","volume":"47","author":"RV Dur\u00e1n","year":"2012","unstructured":"Dur\u00e1n, R. V. et al. Glutaminolysis Activates Rag-mTORC1 Signaling. Mol. Cell 47, 349\u2013358 (2012).","journal-title":"Mol. Cell"},{"key":"BFsrep27698_CR34","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1126\/science.aab2674","volume":"351","author":"RL Wolfson","year":"2016","unstructured":"Wolfson, R. L. et al. Sestrin2 is a leucine sensor for the mTORC1 pathway. Science 351, 43\u201348 (2016).","journal-title":"Science"},{"key":"BFsrep27698_CR35","doi-asserted-by":"publisher","first-page":"2331","DOI":"10.1101\/gad.269324.115","volume":"29","author":"J Ye","year":"2015","unstructured":"Ye, J. et al. GCN2 sustains mTORC1 suppression upon amino acid deprivation by inducing Sestrin2. Genes Dev. 29, 2331\u20132336 (2015).","journal-title":"Genes Dev."},{"key":"BFsrep27698_CR36","doi-asserted-by":"publisher","first-page":"14484","DOI":"10.1074\/jbc.273.23.14484","volume":"273","author":"K Hara","year":"1998","unstructured":"Hara, K. et al. Amino acid sufficiency and mTOR regulate p70 S6 kinase and eIF-4E BP1 through a common effector mechanism. J Biol Chem 273, 14484\u201394 (1998).","journal-title":"J Biol Chem"},{"key":"BFsrep27698_CR37","doi-asserted-by":"publisher","first-page":"786","DOI":"10.1016\/j.cell.2014.01.024","volume":"156","author":"C Demetriades","year":"2014","unstructured":"Demetriades, C., Doumpas, N. & Teleman, A. A. Regulation of TORC1 in Response to Amino Acid Starvation via Lysosomal Recruitment of TSC2. Cell 156, 786\u2013799 (2014).","journal-title":"Cell"},{"key":"BFsrep27698_CR38","doi-asserted-by":"publisher","first-page":"619","DOI":"10.1016\/S1097-2765(03)00105-9","volume":"11","author":"HP Harding","year":"2003","unstructured":"Harding, H. P. et al. An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol. Cell 11, 619\u2013633 (2003).","journal-title":"Mol. Cell"},{"key":"BFsrep27698_CR39","doi-asserted-by":"publisher","first-page":"1165","DOI":"10.1016\/S1097-2765(01)00265-9","volume":"7","author":"D Scheuner","year":"2001","unstructured":"Scheuner, D. et al. Translational Control is Required for the Unfolded Protein Response and In Vivo Glucose Homeostasis. Mol. Cell 7, 1165\u20131176 (2001).","journal-title":"Mol. Cell"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep27698.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep27698","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep27698.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T05:29:50Z","timestamp":1672896590000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep27698"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,6,14]]},"references-count":39,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2016,6,28]]}},"alternative-id":["BFsrep27698"],"URL":"https:\/\/doi.org\/10.1038\/srep27698","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,6,14]]},"assertion":[{"value":"9 December 2015","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 May 2016","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 June 2016","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"27698"}}