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A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling

mTORC1 and GCN2 are serine/threonine kinases that control how cells adapt to amino acid availability. mTORC1 responds to amino acids to promote translation and cell growth while GCN2 senses limiting amino acids to hinder translation via eIF2α phosphorylation. GCN2 is an appealing target for cancer t...

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Autores principales: Brüggenthies, Johanna B., Fiore, Alessandra, Russier, Marion, Bitsina, Christina, Brötzmann, Julian, Kordes, Susanne, Menninger, Sascha, Wolf, Alexander, Conti, Elena, Eickhoff, Jan E., Murray, Peter J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9668732/
https://www.ncbi.nlm.nih.gov/pubmed/36273589
http://dx.doi.org/10.1016/j.jbc.2022.102629
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author Brüggenthies, Johanna B.
Fiore, Alessandra
Russier, Marion
Bitsina, Christina
Brötzmann, Julian
Kordes, Susanne
Menninger, Sascha
Wolf, Alexander
Conti, Elena
Eickhoff, Jan E.
Murray, Peter J.
author_facet Brüggenthies, Johanna B.
Fiore, Alessandra
Russier, Marion
Bitsina, Christina
Brötzmann, Julian
Kordes, Susanne
Menninger, Sascha
Wolf, Alexander
Conti, Elena
Eickhoff, Jan E.
Murray, Peter J.
author_sort Brüggenthies, Johanna B.
collection PubMed
description mTORC1 and GCN2 are serine/threonine kinases that control how cells adapt to amino acid availability. mTORC1 responds to amino acids to promote translation and cell growth while GCN2 senses limiting amino acids to hinder translation via eIF2α phosphorylation. GCN2 is an appealing target for cancer therapies because malignant cells can harness the GCN2 pathway to temper the rate of translation during rapid amino acid consumption. To isolate new GCN2 inhibitors, we created cell-based, amino acid limitation reporters via genetic manipulation of Ddit3 (encoding the transcription factor CHOP). CHOP is strongly induced by limiting amino acids and in this context, GCN2-dependent. Using leucine starvation as a model for essential amino acid sensing, we unexpectedly discovered ATP-competitive PI3 kinase-related kinase inhibitors, including ATR and mTOR inhibitors like torins, completely reversed GCN2 activation in a time-dependent way. Mechanistically, via inhibiting mTORC1-dependent translation, torins increased intracellular leucine, which was sufficient to reverse GCN2 activation and the downstream integrated stress response including stress-induced transcriptional factor ATF4 expression. Strikingly, we found that general translation inhibitors mirrored the effects of torins. Therefore, we propose that mTOR kinase inhibitors concurrently inhibit different branches of amino acid sensing by a dual mechanism involving direct inhibition of mTOR and indirect suppression of GCN2 that are connected by effects on the translation machinery. Collectively, our results highlight distinct ways of regulating GCN2 activity.
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spelling pubmed-96687322022-11-17 A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling Brüggenthies, Johanna B. Fiore, Alessandra Russier, Marion Bitsina, Christina Brötzmann, Julian Kordes, Susanne Menninger, Sascha Wolf, Alexander Conti, Elena Eickhoff, Jan E. Murray, Peter J. J Biol Chem Research Article mTORC1 and GCN2 are serine/threonine kinases that control how cells adapt to amino acid availability. mTORC1 responds to amino acids to promote translation and cell growth while GCN2 senses limiting amino acids to hinder translation via eIF2α phosphorylation. GCN2 is an appealing target for cancer therapies because malignant cells can harness the GCN2 pathway to temper the rate of translation during rapid amino acid consumption. To isolate new GCN2 inhibitors, we created cell-based, amino acid limitation reporters via genetic manipulation of Ddit3 (encoding the transcription factor CHOP). CHOP is strongly induced by limiting amino acids and in this context, GCN2-dependent. Using leucine starvation as a model for essential amino acid sensing, we unexpectedly discovered ATP-competitive PI3 kinase-related kinase inhibitors, including ATR and mTOR inhibitors like torins, completely reversed GCN2 activation in a time-dependent way. Mechanistically, via inhibiting mTORC1-dependent translation, torins increased intracellular leucine, which was sufficient to reverse GCN2 activation and the downstream integrated stress response including stress-induced transcriptional factor ATF4 expression. Strikingly, we found that general translation inhibitors mirrored the effects of torins. Therefore, we propose that mTOR kinase inhibitors concurrently inhibit different branches of amino acid sensing by a dual mechanism involving direct inhibition of mTOR and indirect suppression of GCN2 that are connected by effects on the translation machinery. Collectively, our results highlight distinct ways of regulating GCN2 activity. American Society for Biochemistry and Molecular Biology 2022-10-20 /pmc/articles/PMC9668732/ /pubmed/36273589 http://dx.doi.org/10.1016/j.jbc.2022.102629 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Brüggenthies, Johanna B.
Fiore, Alessandra
Russier, Marion
Bitsina, Christina
Brötzmann, Julian
Kordes, Susanne
Menninger, Sascha
Wolf, Alexander
Conti, Elena
Eickhoff, Jan E.
Murray, Peter J.
A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling
title A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling
title_full A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling
title_fullStr A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling
title_full_unstemmed A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling
title_short A cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase GCN2 signaling
title_sort cell-based chemical-genetic screen for amino acid stress response inhibitors reveals torins reverse stress kinase gcn2 signaling
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9668732/
https://www.ncbi.nlm.nih.gov/pubmed/36273589
http://dx.doi.org/10.1016/j.jbc.2022.102629
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