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Modulation of glutamine metabolism by the PI(3)K–PKB–FOXO network regulates autophagy

Nature Cell Biology volume 14, pages 829–837 (2012)Cite this article

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Abstract

The PI(3)K–PKB–FOXO signalling network provides a major intracellular hub for the regulation of cell proliferation, survival and stress resistance. Here we report an unexpected role for FOXO transcription factors in regulating autophagy by modulating intracellular glutamine levels. To identify transcriptional targets of this network, we performed global transcriptional analyses after conditional activation of the key components PI(3)K, PKB/Akt, FOXO3 and FOXO4. Using this pathway approach, we identified glutamine synthetase as being transcriptionally regulated by PI(3)K–PKB–FOXO signalling. Conditional activation of FOXO also led to an increased level of glutamine production. FOXO activation resulted in mTOR inhibition by preventing the translocation of mTOR to lysosomal membranes in a glutamine-synthetase-dependent manner. This resulted in an increased level of autophagy as measured by LC3 lipidation, p62 degradation and fluorescent imaging of multiple autophagosomal markers. Inhibition of FOXO3-mediated autophagy increased the level of apoptosis, suggesting that the induction of autophagy by FOXO3-mediated glutamine synthetase expression is important for cellular survival. These findings reveal a growth-factor-responsive network that can directly modulate autophagy through the regulation of glutamine metabolism.

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Figure 1: Modulation of the PI(3)K–PKB–FOXO signalling network results in regulation of glutamine synthetase mRNA and protein levels.
Figure 2: FOXO-mediated regulation of glutamine synthetase expression is transcriptionally regulated and evolutionarily conserved.
Figure 3: Modulation of the PI(3)K–PKB–FOXO pathway results in specific regulation of glutamine levels.
Figure 4: Regulation of glutamine synthetase expression by PI(3)K–PKB–FOXO inhibits mTOR and modulates autophagy.
Figure 5: FOXO-induced glutamine synthetase expression increases the level of autophagy.
Figure 6: FOXO-mediated glutamine synthetase activation induces autophagosome formation and increases the level of survival.

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Acknowledgements

We thank R. Korswagen for helpful discussions. K.E.v.d.V. was supported by a grant from the Dutch Scientific Organisation (NWO-VIDI), R.G. was supported by grants (0313081F and 0315735, Virtual Liver) from the German Federal Ministry of Education and Research (BMBF), T.P-C. was supported by the German Research Foundation (DFG; SFB 773, A03), R.v.B. was supported by a grant from CTMM and L.P.V. was supported by a grant from NIRM.

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Author notes

  1. Kristan E. van der Vos

    Present address: Present address: Departments of Neurology and Radiology, Massachusetts General Hospital, and Neuroscience Program, Harvard Medical School, Boston, Massachusetts 02129, USA,

  2. Pernilla Eliasson and Tassula Proikas-Cezanne: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Immunology, University Medical Center Utrecht, Utrecht 3584CX, The Netherlands

    Kristan E. van der Vos & Paul J. Coffer

  2. Department of Cell Biology, University Medical Center Utrecht, Utrecht 3584CX, The Netherlands

    Pernilla Eliasson, Stephin J. Vervoort, Ruben van Boxtel, Cornelieke Pals, Liesbeth P. Verhagen, A. Koen Braat & Paul J. Coffer

  3. Autophagy Laboratory, Interfaculty Institute for Cell Biology, University of Tuebingen, Tuebingen 72076, Germany

    Tassula Proikas-Cezanne & Mario Mauthe

  4. Department of Molecular Cancer Research, University Medical Center, Utrecht 3584CX, The Netherlands

    Marrit Putker, Iris J. van Zutphen, Marian J. A. Groot Koerkamp, Tobias B. Dansen, Frank C. Holstege & Boudewijn M. Burgering

  5. Institut für Biochemie, Medizinische Fakultät, Universität Leipzig, Leipzig 04103, Germany

    Sebastian Zellmer & Rolf Gebhardt

  6. Center for Molecular and Cellular Intervention, Wilhelmina Children’s Hospital, Utrecht 3584EA, The Netherlands

    Stephin J. Vervoort, A. Koen Braat & Paul J. Coffer

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Contributions

K.E.v.d.V. was involved in experimental strategy and design, performed experiments, analysed data and wrote the paper. P.E., S.J.V., R.v.B., M.P., I.J.v.Z., M.M., S.Z., C.P., L.P.V., M.J.A.G.K., A.K.B. and T.B.D. performed experiments. T.P-C., F.C.H., R.G. and B.M.B. were involved in experimental design. P.J.C. was involved in experimental strategy and design and wrote the paper.

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Correspondence to Paul J. Coffer.

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van der Vos, K., Eliasson, P., Proikas-Cezanne, T. et al. Modulation of glutamine metabolism by the PI(3)K–PKB–FOXO network regulates autophagy. Nat Cell Biol 14, 829–837 (2012). https://doi.org/10.1038/ncb2536

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