Abstract

The major energy source for most cells is glucose, from which ATP is generated via glycolysis and/or oxidative metabolism. Glucose deprivation activates AMP-activated protein kinase (AMPK)1, but it is unclear whether this activation occurs solely via changes in AMP or ADP, the classical activators of AMPK2,3,4,5. Here, we describe an AMP/ADP-independent mechanism that triggers AMPK activation by sensing the absence of fructose-1,6-bisphosphate (FBP), with AMPK being progressively activated as extracellular glucose and intracellular FBP decrease. When unoccupied by FBP, aldolases promote the formation of a lysosomal complex containing at least v-ATPase, ragulator, axin, liver kinase B1 (LKB1) and AMPK, which has previously been shown to be required for AMPK activation6,7. Knockdown of aldolases activates AMPK even in cells with abundant glucose, whereas the catalysis-defective D34S aldolase mutant, which still binds FBP, blocks AMPK activation. Cell-free reconstitution assays show that addition of FBP disrupts the association of axin and LKB1 with v-ATPase and ragulator. Importantly, in some cell types AMP/ATP and ADP/ATP ratios remain unchanged during acute glucose starvation, and intact AMP-binding sites on AMPK are not required for AMPK activation. These results establish that aldolase, as well as being a glycolytic enzyme, is a sensor of glucose availability that regulates AMPK.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , & AMP-activated protein kinase is activated by low glucose in cell lines derived from pancreatic beta cells, and may regulate insulin release. Biochem. J. 335, 533–539 (1998)

  2. 2.

    , & AMP-activated protein kinase: a cellular energy sensor that comes in 12 flavours. FEBS J. 283, 2987–3001 (2016)

  3. 3.

    AMPK signalling in health and disease. Curr. Opin. Cell Biol. 45, 31–37 (2017)

  4. 4.

    & AMPK in health and disease. Physiol. Rev. 89, 1025–1078 (2009)

  5. 5.

    & The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat. Cell Biol. 13, 1016–1023 (2011)

  6. 6.

    et al. The lysosomal v-ATPase-Ragulator complex is a common activator for AMPK and mTORC1, acting as a switch between catabolism and anabolism. Cell Metab. 20, 526–540 (2014)

  7. 7.

    et al. AMP as a low-energy charge signal autonomously initiates assembly of AXIN-AMPK-LKB1 complex for AMPK activation. Cell Metab. 18, 546–555 (2013)

  8. 8.

    et al. β-Subunit myristoylation is the gatekeeper for initiating metabolic stress sensing by AMP-activated protein kinase (AMPK). Proc. Natl Acad. Sci. USA 107, 19237–19241 (2010)

  9. 9.

    et al. Use of cells expressing gamma subunit variants to identify diverse mechanisms of AMPK activation. Cell Metab. 11, 554–565 (2010)

  10. 10.

    & Lysosomes and autophagy in cell death control. Nat. Rev. Cancer 5, 886–897 (2005)

  11. 11.

    & Choreography of AMPK activation. Cell Res. 25, 5–6 (2015)

  12. 12.

    et al. Structural basis of AMPK regulation by small molecule activators. Nat. Commun. 4, 3017 (2013)

  13. 13.

    et al. Investigation of LKB1 Ser431 phosphorylation and Cys433 farnesylation using mouse knockin analysis reveals an unexpected role of prenylation in regulating AMPK activity. Biochem. J. 458, 41–56 (2014)

  14. 14.

    , , & Influence of phosphorylation on the interaction of effectors with rat liver pyruvate kinase. J. Biol. Chem. 257, 233–240 (1982)

  15. 15.

    & Targeting of several glycolytic enzymes using RNA interference reveals aldolase affects cancer cell proliferation through a non-glycolytic mechanism. J. Biol. Chem. 287, 42554–42563 (2012)

  16. 16.

    & Site-directed mutagenesis identifies aspartate 33 as a previously unidentified critical residue in the catalytic mechanism of rabbit aldolase A. J. Biol. Chem. 268, 1095–1100 (1993)

  17. 17.

    , , , & Snapshots of catalysis: the structure of fructose-1,6-(bis)phosphate aldolase covalently bound to the substrate dihydroxyacetone phosphate. Biochemistry 40, 13868–13875 (2001)

  18. 18.

    & The complete amino acid sequence for the anaerobically induced aldolase from maize derived from cDNA clones. Plant Physiol. 82, 1076–1080 (1986)

  19. 19.

    , , & Enzyme substrate specificity conferred by distinct conformational pathways. J. Am. Chem. Soc. 137, 13876–13886 (2015)

  20. 20.

    Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology. Nat. Rev. Mol. Cell Biol. 8, 917–929 (2007)

  21. 21.

    , , , & Interaction between aldolase and vacuolar H+-ATPase: evidence for direct coupling of glycolysis to the ATP-hydrolyzing proton pump. J. Biol. Chem. 276, 30407–30413 (2001)

  22. 22.

    , , & The glycolytic enzyme aldolase mediates assembly, expression, and activity of vacuolar H+-ATPase. J. Biol. Chem. 279, 8732–8739 (2004)

  23. 23.

    , , , & Physical interaction between aldolase and vacuolar H+-ATPase is essential for the assembly and activity of the proton pump. J. Biol. Chem. 282, 24495–24503 (2007)

  24. 24.

    et al. Structure of mammalian AMPK and its regulation by ADP. Nature 472, 230–233 (2011)

  25. 25.

    , & Glucose repression/derepression in budding yeast: SNF1 protein kinase is activated by phosphorylation under derepressing conditions, and this correlates with a high AMP:ATP ratio. Curr. Biol. 6, 1426–1434 (1996)

  26. 26.

    et al. ULK1/2 constitute a bifurcate node controlling glucose metabolic fluxes in addition to autophagy. Mol. Cell 62, 359–370 (2016)

  27. 27.

    et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 29, 341–345 (2011)

  28. 28.

    et al. Genome engineering using the CRISPR-Cas9 system. Nat. Protocols 8, 2281–2308 (2013)

  29. 29.

    , , , & Aldolase mediates the association of F-actin with the insulin-responsive glucose transporter GLUT4. J. Biol. Chem. 274, 17742–17747 (1999)

  30. 30.

    et al. Automatic and quantitative measurement of protein-protein colocalization in live cells. Biophys. J. 86, 3993–4003 (2004)

  31. 31.

    et al. Phosphoinositide 3-kinase regulates glycolysis through mobilization of aldolase from the actin cytoskeleton. Cell 164, 433–446 (2016)

  32. 32.

    et al. Study of polar metabolites in tobacco from different geographical origins by using capillary electrophoresis–mass spectrometry. Metabolomics 10, (2014)

  33. 33.

    et al. A metabolomics study delineating geographical location-associated primary metabolic changes in the leaves of growing tobacco plants by GC-MS and CE-MS. Sci. Rep. 5, 16346 (2015)

  34. 34.

    & in Current Topics in Cellular Regulation Vol. 2 (eds Horecker, B. L. & Stadtman, E. R.) 227–273 (Academic, 1970)

  35. 35.

    , & Metabolism, cell surface organization, and disease. Cell 139, 1229–1241 (2009)

Download references

Acknowledgements

We thank all other members of the S.-C.L. laboratory for suggestions and technical assistance, and the proteomics team at the University of Dundee (D. Lamont, A. Atrih, W. Chen and K. Beattie) for LC–MS analyses of nucleotides. D.G.H. was supported by an Investigator Award from the Wellcome Trust (097726) and a Programme Grant from Cancer Research UK (C37030/A15101). S.-C.L. was supported by grants from the National Key Research and Development Project of China (2016YFA0502001) and the National Natural Science Foundation of China (#31430094, #31690101, #31571214, #31601152 and #J1310027).

Author information

Author notes

    • Chen-Song Zhang
    • , Simon A. Hawley
    • , Yue Zong
    •  & Mengqi Li

    These authors contributed equally to this work.

Affiliations

  1. State Key Laboratory for Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Fujian 361102, China

    • Chen-Song Zhang
    • , Yue Zong
    • , Mengqi Li
    • , Teng Ma
    • , Jiwen Cui
    • , Jin-Wei Feng
    • , Yu-Qing Wu
    • , Terytty Yang Li
    • , Zhiyun Ye
    • , Shu-Yong Lin
    •  & Sheng-Cai Lin
  2. Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK

    • Simon A. Hawley
    • , Alexander Gray
    •  & D. Grahame Hardie
  3. Scientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Liaoning 116023, China

    • Zhichao Wang
    •  & Hai-Long Piao
  4. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Liaoning 116023, China

    • Zhichao Wang
  5. University of Chinese Academy of Sciences, Beijing 100049, China

    • Zhichao Wang
  6. Key Laboratory of Food Safety Research, Institute for Nutritional Sciences (INS), Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS), Shanghai 200031, China

    • Mingjiang Zhu
    •  & Huiyong Yin

Authors

  1. Search for Chen-Song Zhang in:

  2. Search for Simon A. Hawley in:

  3. Search for Yue Zong in:

  4. Search for Mengqi Li in:

  5. Search for Zhichao Wang in:

  6. Search for Alexander Gray in:

  7. Search for Teng Ma in:

  8. Search for Jiwen Cui in:

  9. Search for Jin-Wei Feng in:

  10. Search for Mingjiang Zhu in:

  11. Search for Yu-Qing Wu in:

  12. Search for Terytty Yang Li in:

  13. Search for Zhiyun Ye in:

  14. Search for Shu-Yong Lin in:

  15. Search for Huiyong Yin in:

  16. Search for Hai-Long Piao in:

  17. Search for D. Grahame Hardie in:

  18. Search for Sheng-Cai Lin in:

Contributions

C.-S.Z., S.A.H., Y.Z., M.L., D.G.H. and S.-C.L. conceived the study. C.-S.Z., S.A.H., Y.Z. and M.L. performed most experiments with assistance from T.M., J.C., J.-W.F., A.G., Y.-Q.W., T.Y.L. and S.-Y.L. Y.Z. and Z.W. performed the CE–MS-based analysis of metabolites, and S.A.H., M.L. and M.Z. performed the analysis of adenylates. Z.Y., S.-Y.L., H.Y. and H.-L.P. helped with discussion and interpretation of results. D.G.H. and S.-C.L. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to D. Grahame Hardie or Sheng-Cai Lin.

Reviewer Information Nature thanks D. Tolan and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Notes 1-3 and the uncropped gels.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature23275

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.