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Inactivation of Rheb by PRAK-mediated phosphorylation is essential for energy-depletion-induced suppression of mTORC1

Nature Cell Biology volume 13, pages 263–272 (2011)Cite this article

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Abstract

Cell growth can be suppressed by stressful environments, but the role of stress pathways in this process is largely unknown. Here we show that a cascade of p38β mitogen-activated protein kinase (MAPK) and p38-regulated/activated kinase (PRAK) plays a role in energy-starvation-induced suppression of mammalian target of rapamycin (mTOR), and that energy starvation activates the p38β–PRAK cascade. Depletion of p38β or PRAK diminishes the suppression of mTOR complex 1 (mTORC1) and reduction of cell size induced by energy starvation. We show that p38β–PRAK operates independently of the known mTORC1 inactivation pathways—phosphorylation of tuberous sclerosis protein 2 (TSC2) and Raptor by AMP-activated protein kinase (AMPK)—and surprisingly, that PRAK directly regulates Ras homologue enriched in brain (Rheb), a key component of the mTORC1 pathway, by phosphorylation. Phosphorylation of Rheb at Ser 130 by PRAK impairs the nucleotide-binding ability of Rheb and inhibits Rheb-mediated mTORC1 activation. The direct regulation of Rheb by PRAK integrates a stress pathway with the mTORC1 pathway in response to energy depletion.

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Figure 1: p38β is essential for the 2-DG-induced cellular response.
Figure 2: PRAK is essential for the 2-DG-induced cellular response.
Figure 3: p38β and PRAK selectively participate in certain forms of energy-depletion-induced mTORC1 inactivation.
Figure 4: Suppression of mTORC1 by PRAK is independent of AMPK.
Figure 5: Suppression of mTORC1 by PRAK is independent of TSC2.
Figure 6: PRAK regulates Rheb by phosphorylation of Rheb on Ser 130.
Figure 7: Rheb phosphorylation by PRAK reduces the nucleotide binding ability of Rheb.

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Acknowledgements

We would like to thank H. Jiang (Boehringer Ingelheim Pharmaceuticals) for providing p38 conditional knockout mice, B. Viollet (Universite Paris Descartes) and K. R. Laderoute (Stanford University School of Medicine) for A M P K α 1/α 2−/− cells. This work was supported by grants from NSF China 30830092, 30921005, 30828219, 973 program 2009CB22200, National Institutes of Health AI41637 and AI68896.

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Authors and Affiliations

  1. The Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China

    Min Zheng, Yan-Hai Wang, Xiao-Nan Wu, Su-Qin Wu, Sheng-Cai Lin & Jiahuai Han

  2. The National Institute of Biological Sciences, Beijing 102206, China

    Bao-Ju Lu & Meng-Qiu Dong

  3. Department of Physiology, National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China

    Hongbing Zhang

  4. The Scripps Research Institute, La Jolla, California 92037, USA

    Peiqing Sun

  5. Department of Pharmacology and Moores Cancer Center, University of California, San Diego, La Jolla, California 92093-0815, USA

    Kun-Liang Guan

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Contributions

M.Z., Y-H.W., X-N.W., S-Q.W. and J.H. designed and carried out the experiments. B-J.L. and M-Q.D. carried out mass spectrometry analysis. M.Z., H.Z., P.S., S-C.L., K-L.G. and J.H. participated in the interpretation of the data. M.Z. and J.H. wrote the manuscript.

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Correspondence to Jiahuai Han.

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Zheng, M., Wang, YH., Wu, XN. et al. Inactivation of Rheb by PRAK-mediated phosphorylation is essential for energy-depletion-induced suppression of mTORC1. Nat Cell Biol 13, 263–272 (2011). https://doi.org/10.1038/ncb2168

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