Glutamine and asparagine activate mTORC1 independently of Rag GTPases

Nutrient sensing by cells is crucial, and when this sensing mechanism is disturbed, human disease can occur. mTOR complex 1 (mTORC1) senses amino acids to control cell growth, metabolism, and autophagy. Leucine, arginine, and methionine signal to mTORC1 through the well-characterized Rag GTPase signaling pathway. In contrast, glutamine activates mTORC1 through a Rag GTPase–independent mechanism that requires ADP-ribosylation factor 1 (Arf1). Here, using several biochemical and genetic approaches, we show that eight amino acids filter through the Rag GTPase pathway. Like glutamine, asparagine signals to mTORC1 through Arf1 in the absence of the Rag GTPases. Both the Rag-dependent and Rag-independent pathways required the lysosome and lysosomal function for mTORC1 activation. Our results show that mTORC1 is differentially regulated by amino acids through two distinct pathways.

[1]  Abdullahi Umar Ibrahim,et al.  Genome Engineering Using the CRISPR Cas9 System , 2019 .

[2]  Michael D. Brooks,et al.  Asparagine and Glutamine: Co-conspirators Fueling Metastasis. , 2018, Cell metabolism.

[3]  T. Noda,et al.  Gtr/Ego-independent TORC1 activation is achieved through a glutamine-sensitive interaction with Pib2 on the vacuolar membrane , 2018, PLoS genetics.

[4]  R. White,et al.  As Extracellular Glutamine Levels Decline, Asparagine Becomes an Essential Amino Acid. , 2018, Cell metabolism.

[5]  N. Pavletich,et al.  Structural Mechanisms of mTORC1 activation by RHEB and inhibition by PRAS40 , 2017, Nature.

[6]  S. Gygi,et al.  SAMTOR is an S-adenosylmethionine sensor for the mTORC1 pathway , 2017, Science.

[7]  Gregory A. Wyant,et al.  Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes , 2017, Science.

[8]  Gregory A. Wyant,et al.  mTORC1 Activator SLC38A9 Is Required to Efflux Essential Amino Acids from Lysosomes and Use Protein as a Nutrient , 2017, Cell.

[9]  C. Thompson,et al.  Nutrient acquisition strategies of mammalian cells , 2017, Nature.

[10]  T. Maeda,et al.  An In Vitro TORC1 Kinase Assay That Recapitulates the Gtr-Independent Glutamine-Responsive TORC1 Activation Mechanism on Yeast Vacuoles , 2017, Molecular and Cellular Biology.

[11]  David M. Sabatini,et al.  mTOR Signaling in Growth, Metabolism, and Disease , 2017, Cell.

[12]  D. Sabatini,et al.  mTOR Signaling in Growth, Metabolism, and Disease , 2017, Cell.

[13]  Gregory A. Wyant,et al.  KICSTOR recruits GATOR1 to the lysosome and is necessary for nutrients to regulate mTORC1 , 2017, Nature.

[14]  D. Sabatini,et al.  Mechanism of arginine sensing by CASTOR1 upstream of mTORC1 , 2016, Nature.

[15]  T. Graeber,et al.  Asparagine promotes cancer cell proliferation through use as an amino acid exchange factor , 2016, Nature Communications.

[16]  Gregory A. Wyant,et al.  The CASTOR Proteins Are Arginine Sensors for the mTORC1 Pathway , 2016, Cell.

[17]  D. Sabatini,et al.  Sestrin2 is a leucine sensor for the mTORC1 pathway , 2016, Science.

[18]  D. Sabatini,et al.  Structural basis for leucine sensing by the Sestrin2-mTORC1 pathway , 2016, Science.

[19]  K. Cunningham,et al.  A LAPF/phafin1-like protein regulates TORC1 and lysosomal membrane permeabilization in response to endoplasmic reticulum membrane stress , 2015, Molecular biology of the cell.

[20]  J. Blenis,et al.  A nexus for cellular homeostasis: the interplay between metabolic and signal transduction pathways. , 2015, Current opinion in biotechnology.

[21]  C. Behrends,et al.  Amino Acid-Dependent mTORC1 Regulation by the Lysosomal Membrane Protein SLC38A9 , 2015, Molecular and Cellular Biology.

[22]  Gregory A. Wyant,et al.  Lysosomal amino acid transporter SLC38A9 signals arginine sufficiency to mTORC1 , 2015, Science.

[23]  K. Guan,et al.  Differential regulation of mTORC1 by leucine and glutamine , 2015, Science.

[24]  G. Superti-Furga,et al.  SLC38A9 is a component of the lysosomal amino acid-sensing machinery that controls mTORC1 , 2014, Nature.

[25]  Steven P Gygi,et al.  The Sestrins interact with GATOR2 to negatively regulate the amino-acid-sensing pathway upstream of mTORC1. , 2014, Cell reports.

[26]  Florian Rudroff,et al.  Nitrogen Source Activates TOR (Target of Rapamycin) Complex 1 via Glutamine and Independently of Gtr/Rag Proteins* , 2014, The Journal of Biological Chemistry.

[27]  L. Cantley,et al.  Spatial Control of the TSC Complex Integrates Insulin and Nutrient Regulation of mTORC1 at the Lysosome , 2014, Cell.

[28]  David A. Scott,et al.  Genome engineering using the CRISPR-Cas9 system , 2013, Nature Protocols.

[29]  D. Sabatini,et al.  The folliculin tumor suppressor is a GAP for the RagC/D GTPases that signal amino acid levels to mTORC1. , 2013, Molecular cell.

[30]  K. Guan,et al.  Nutrient signaling to mTOR and cell growth. , 2013, Trends in biochemical sciences.

[31]  Matthew Meyerson,et al.  A Tumor Suppressor Complex with GAP Activity for the Rag GTPases That Signal Amino Acid Sufficiency to mTORC1 , 2013, Science.

[32]  K. Guan,et al.  Amino acid signalling upstream of mTOR , 2013, Nature Reviews Molecular Cell Biology.

[33]  Sang Gyun Kim,et al.  Metabolic stress controls mTORC1 lysosomal localization and dimerization by regulating the TTT-RUVBL1/2 complex. , 2013, Molecular cell.

[34]  D. Sabatini,et al.  Ragulator Is a GEF for the Rag GTPases that Signal Amino Acid Levels to mTORC1 , 2012, Cell.

[35]  E. Gottlieb,et al.  Glutaminolysis activates Rag-mTORC1 signaling. , 2012, Molecular cell.

[36]  M. G. Koerkamp,et al.  Modulation of glutamine metabolism by the PI(3)K–PKB–FOXO network regulates autophagy , 2012, Nature Cell Biology.

[37]  A. Ballabio,et al.  A lysosome-to-nucleus signalling mechanism senses and regulates the lysosome via mTOR and TFEB , 2012, The EMBO journal.

[38]  Roberto Zoncu,et al.  mTORC1 Senses Lysosomal Amino Acids Through an Inside-Out Mechanism That Requires the Vacuolar H+-ATPase , 2011, Science.

[39]  K. Guan,et al.  Crystal structure of the Gtr1p-Gtr2p complex reveals new insights into the amino acid-induced TORC1 activation. , 2011, Genes & development.

[40]  D. Sabatini,et al.  mTOR: from growth signal integration to cancer, diabetes and ageing , 2010, Nature Reviews Molecular Cell Biology.

[41]  D. Burrin,et al.  Arginine-induced stimulation of protein synthesis and survival in IPEC-J2 cells is mediated by mTOR but not nitric oxide. , 2010, American journal of physiology. Endocrinology and metabolism.

[42]  D. Sabatini,et al.  Ragulator-Rag Complex Targets mTORC1 to the Lysosomal Surface and Is Necessary for Its Activation by Amino Acids , 2010, Cell.

[43]  K. Inoki,et al.  Tuberous sclerosis complex, implication from a rare genetic disease to common cancer treatment. , 2009, Human molecular genetics.

[44]  Jeffrey P. MacKeigan,et al.  Bidirectional Transport of Amino Acids Regulates mTOR and Autophagy , 2009, Cell.

[45]  T. P. Neufeld,et al.  Regulation of TORC1 by Rag GTPases in nutrient response , 2008, Nature Cell Biology.

[46]  David M. Sabatini,et al.  The Rag GTPases Bind Raptor and Mediate Amino Acid Signaling to mTORC1 , 2008, Science.

[47]  Michael Forgac,et al.  Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology , 2007, Nature Reviews Molecular Cell Biology.

[48]  D. Kwiatkowski,et al.  Tuberous sclerosis: a GAP at the crossroads of multiple signaling pathways. , 2005, Human molecular genetics.

[49]  J. Blenis,et al.  Tuberous Sclerosis Complex Gene Products, Tuberin and Hamartin, Control mTOR Signaling by Acting as a GTPase-Activating Protein Complex toward Rheb , 2003, Current Biology.

[50]  K. Inoki,et al.  Rheb GTPase is a direct target of TSC2 GAP activity and regulates mTOR signaling. , 2003, Genes & development.

[51]  B. Edgar,et al.  Rheb is a direct target of the tuberous sclerosis tumour suppressor proteins , 2003, Nature Cell Biology.

[52]  T. Nishi,et al.  The vacuolar (H+)-ATPases — nature's most versatile proton pumps , 2002, Nature Reviews Molecular Cell Biology.

[53]  E. Henske,et al.  Mutations in the tuberous sclerosis complex gene TSC2 are a cause of sporadic pulmonary lymphangioleiomyomatosis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[54]  J. Avruch,et al.  Amino Acid Sufficiency and mTOR Regulate p70 S6 Kinase and eIF-4E BP1 through a Common Effector Mechanism* , 1998, The Journal of Biological Chemistry.

[55]  J. Rothman,et al.  Inhibition by brefeldin A of a Golgi membrane enzyme that catalyses exchange of guanine nucleotide bound to ARF , 1992, Nature.

[56]  E. Frame.  The levels of individual free amino acids in the plasma of normal man at various intervals after a high-protein meal. , 1958, The Journal of clinical investigation.

[57]  S. Moore,et al.  The free amino acids of human blood plasma. , 1954, The Journal of biological chemistry.