The E3 ubiquitin ligase ZNRF2 is a substrate of mTORC1 and regulates its activation by amino acids

Gerta Hoxhaj (Lead / Corresponding author), Edward Caddye, Ayaz Najafov, Vanessa P. Houde, Catherine Johnson, Kumara Dissanayake, Rachel Toth, David G. Campbell, Alan R. Prescott, Carol MacKintosh (Lead / Corresponding author)

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Abstract

The mechanistic Target of Rapamycin complex 1 (mTORC1) senses intracellular amino acid levels through an intricate machinery, which includes the Rag GTPases, Ragulator and vacuolar ATPase (V-ATPase). The membrane-associated E3 ubiquitin ligase ZNRF2 is released into the cytosol upon its phosphorylation by Akt. In this study, we show that ZNRF2 interacts with mTOR on membranes, promoting the amino acid-stimulated translocation of mTORC1 to lysosomes and its activation in human cells. ZNRF2 also interacts with the V-ATPase and preserves lysosomal acidity. Moreover, knockdown of ZNRF2 decreases cell size and cell proliferation. Upon growth factor and amino acid stimulation, mTORC1 phosphorylates ZNRF2 on Ser145, and this phosphosite is dephosphorylated by protein phosphatase 6. Ser145 phosphorylation stimulates vesicle-to-cytosol translocation of ZNRF2 and forms a novel negative feedback on mTORC1. Our findings uncover ZNRF2 as a component of the amino acid sensing machinery that acts upstream of Rag-GTPases and the V-ATPase to activate mTORC1.

Original languageEnglish
Article numbere12278
Pages (from-to)1-18
Number of pages18
JournaleLife
Volume5
DOIs
Publication statusPublished - 22 Apr 2016

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Aminoacylation
Ubiquitin-Protein Ligases
Vacuolar Proton-Translocating ATPases
Chemical activation
Amino Acids
Substrates
Phosphorylation
GTP Phosphohydrolases
Cytosol
Machinery
Membranes
Cell proliferation
Lysosomes
Cell Size
Acidity
Intercellular Signaling Peptides and Proteins
Cells
Cell Proliferation
mechanistic target of rapamycin complex 1
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title = "The E3 ubiquitin ligase ZNRF2 is a substrate of mTORC1 and regulates its activation by amino acids",
abstract = "The mechanistic Target of Rapamycin complex 1 (mTORC1) senses intracellular amino acid levels through an intricate machinery, which includes the Rag GTPases, Ragulator and vacuolar ATPase (V-ATPase). The membrane-associated E3 ubiquitin ligase ZNRF2 is released into the cytosol upon its phosphorylation by Akt. In this study, we show that ZNRF2 interacts with mTOR on membranes, promoting the amino acid-stimulated translocation of mTORC1 to lysosomes and its activation in human cells. ZNRF2 also interacts with the V-ATPase and preserves lysosomal acidity. Moreover, knockdown of ZNRF2 decreases cell size and cell proliferation. Upon growth factor and amino acid stimulation, mTORC1 phosphorylates ZNRF2 on Ser145, and this phosphosite is dephosphorylated by protein phosphatase 6. Ser145 phosphorylation stimulates vesicle-to-cytosol translocation of ZNRF2 and forms a novel negative feedback on mTORC1. Our findings uncover ZNRF2 as a component of the amino acid sensing machinery that acts upstream of Rag-GTPases and the V-ATPase to activate mTORC1.",
author = "Gerta Hoxhaj and Edward Caddye and Ayaz Najafov and Houde, {Vanessa P.} and Catherine Johnson and Kumara Dissanayake and Rachel Toth and Campbell, {David G.} and Prescott, {Alan R.} and Carol MacKintosh",
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The E3 ubiquitin ligase ZNRF2 is a substrate of mTORC1 and regulates its activation by amino acids. / Hoxhaj, Gerta (Lead / Corresponding author); Caddye, Edward; Najafov, Ayaz; Houde, Vanessa P.; Johnson, Catherine; Dissanayake, Kumara; Toth, Rachel; Campbell, David G.; Prescott, Alan R.; MacKintosh, Carol (Lead / Corresponding author).

In: eLife, Vol. 5, e12278, 22.04.2016, p. 1-18.

Research output: Contribution to journalArticle

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AU - Hoxhaj, Gerta

AU - Caddye, Edward

AU - Najafov, Ayaz

AU - Houde, Vanessa P.

AU - Johnson, Catherine

AU - Dissanayake, Kumara

AU - Toth, Rachel

AU - Campbell, David G.

AU - Prescott, Alan R.

AU - MacKintosh, Carol

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AB - The mechanistic Target of Rapamycin complex 1 (mTORC1) senses intracellular amino acid levels through an intricate machinery, which includes the Rag GTPases, Ragulator and vacuolar ATPase (V-ATPase). The membrane-associated E3 ubiquitin ligase ZNRF2 is released into the cytosol upon its phosphorylation by Akt. In this study, we show that ZNRF2 interacts with mTOR on membranes, promoting the amino acid-stimulated translocation of mTORC1 to lysosomes and its activation in human cells. ZNRF2 also interacts with the V-ATPase and preserves lysosomal acidity. Moreover, knockdown of ZNRF2 decreases cell size and cell proliferation. Upon growth factor and amino acid stimulation, mTORC1 phosphorylates ZNRF2 on Ser145, and this phosphosite is dephosphorylated by protein phosphatase 6. Ser145 phosphorylation stimulates vesicle-to-cytosol translocation of ZNRF2 and forms a novel negative feedback on mTORC1. Our findings uncover ZNRF2 as a component of the amino acid sensing machinery that acts upstream of Rag-GTPases and the V-ATPase to activate mTORC1.

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