Metabolism and ageing are intimately linked. Compared with ad libitum feeding, dietary restriction consistently extends lifespan and delays age-related diseases in evolutionarily diverse organisms1,2. Similar conditions of nutrient limitation and genetic or pharmacological perturbations of nutrient or energy metabolism also have longevity benefits3,4. Recently, several metabolites have been identified that modulate ageing5,6; however, the molecular mechanisms underlying this are largely undefined. Here we show that α-ketoglutarate (α-KG), a tricarboxylic acid cycle intermediate, extends the lifespan of adult Caenorhabditis elegans. ATP synthase subunit β is identified as a novel binding protein of α-KG using a small-molecule target identification strategy termed drug affinity responsive target stability (DARTS)7. The ATP synthase, also known as complex V of the mitochondrial electron transport chain, is the main cellular energy-generating machinery and is highly conserved throughout evolution8,9. Although complete loss of mitochondrial function is detrimental, partial suppression of the electron transport chain has been shown to extend C. elegans lifespan10,11,12,13. We show that α-KG inhibits ATP synthase and, similar to ATP synthase knockdown, inhibition by α-KG leads to reduced ATP content, decreased oxygen consumption, and increased autophagy in both C. elegans and mammalian cells. We provide evidence that the lifespan increase by α-KG requires ATP synthase subunit β and is dependent on target of rapamycin (TOR) downstream. Endogenous α-KG levels are increased on starvation and α-KG does not extend the lifespan of dietary-restricted animals, indicating that α-KG is a key metabolite that mediates longevity by dietary restriction. Our analyses uncover new molecular links between a common metabolite, a universal cellular energy generator and dietary restriction in the regulation of organismal lifespan, thus suggesting new strategies for the prevention and treatment of ageing and age-related diseases.

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  1. 1.

    et al. Caloric restriction delays disease onset and mortality in rhesus monkeys. Science 325, 201–204 (2009)

  2. 2.

    et al. Impact of caloric restriction on health and survival in rhesus monkeys from the NIA study. Nature 489, 318–321 (2012)

  3. 3.

    The genetics of ageing. Nature 464, 504–512 (2010)

  4. 4.

    et al. Rapamycin fed late in life extends lifespan in genetically heterogeneous mice. Nature 460, 392–395 (2009)

  5. 5.

    , , & Oxaloacetate supplementation increases lifespan in Caenorhabditis elegans through an AMPK/FOXO-dependent pathway. Aging Cell 8, 765–768 (2009)

  6. 6.

    et al. N-acylethanolamine signalling mediates the effect of diet on lifespan in Caenorhabditis elegans. Nature 473, 226–229 (2011)

  7. 7.

    et al. Target identification using drug affinity responsive target stability (DARTS). Proc. Natl Acad. Sci. USA 106, 21984–21989 (2009)

  8. 8.

    , , & Structure at 2.8 Å resolution of F1-ATPase from bovine heart mitochondria. Nature 370, 621–628 (1994)

  9. 9.

    The ATP synthase—a splendid molecular machine. Annu. Rev. Biochem. 66, 717–749 (1997)

  10. 10.

    , , , & Mitochondrial respiratory chain deficiency in Caenorhabditis elegans results in developmental arrest and increased life span. J. Biol. Chem. 276, 32240–32246 (2001)

  11. 11.

    et al. Rates of behavior and aging specified by mitochondrial function during development. Science 298, 2398–2401 (2002)

  12. 12.

    et al. A systematic RNAi screen identifies a critical role for mitochondria in C. elegans longevity. Nature Genet. 33, 40–48 (2002)

  13. 13.

    & Lifespan regulation by evolutionarily conserved genes essential for viability. PLoS Genet. 3, e56 (2007)

  14. 14.

    & Genetic, behavioral and environmental determinants of male longevity in Caenorhabditis elegans. Genetics 154, 1597–1610 (2000)

  15. 15.

    & Assessing mitochondrial dysfunction in cells. Biochem. J. 435, 297–312 (2011)

  16. 16.

    & The genetics of caloric restriction in Caenorhabditis elegans. Proc. Natl Acad. Sci. USA 95, 13091–13096 (1998)

  17. 17.

    et al. Lifespan extension by conditions that inhibit translation in Caenorhabditis elegans. Aging Cell 6, 95–110 (2007)

  18. 18.

    , , & The TOR pathway comes of age. Biochim. Biophys. Acta 1790, 1067–1074 (2009)

  19. 19.

    , , & Management of cellular energy by the AMP-activated protein kinase system. FEBS Lett. 546, 113–120 (2003)

  20. 20.

    & Different dietary restriction regimens extend lifespan by both independent and overlapping genetic pathways in C. elegans. Aging Cell 8, 113–127 (2009)

  21. 21.

    , & The target of rapamycin pathway antagonizes pha-4/FoxA to control development and aging. Curr. Biol. 18, 1355–1364 (2008)

  22. 22.

    , , , & PHA-4/Foxa mediates diet-restriction-induced longevity of C. elegans. Nature 447, 550–555 (2007)

  23. 23.

    , & TOR signaling in growth and metabolism. Cell 124, 471–484 (2006)

  24. 24.

    et al. Autophagy genes are essential for dauer development and life-span extension in C. elegans. Science 301, 1387–1391 (2003)

  25. 25.

    & Expanding chemical biology of 2-oxoglutarate oxygenases. Nature Chem. Biol. 4, 152–156 (2008)

  26. 26.

    et al. C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell 107, 43–54 (2001)

  27. 27.

    , , , & The HIF-1 hypoxia-inducible factor modulates lifespan in C. elegans. PLoS ONE 4, e6348 (2009)

  28. 28.

    et al. Conservation of the metabolomic response to starvation across two divergent microbes. Proc. Natl Acad. Sci. USA 103, 19302–19307 (2006)

  29. 29.

    , & Metabolites of citric acid cycle, carbohydrate and phosphorus metabolism, and related reactions, redox and phosphorylating states of hepatic tissue, liver mitochondria and cytosol of the pigeon, under normal feeding and natural nocturnal fasting conditions. Comp. Biochem. Physiol. B 73, 957–963 (1982)

  30. 30.

    et al. Metabolomics approach for analyzing the effects of exercise in subjects with type 1 diabetes mellitus. PLoS ONE 7, e40600 (2012)

  31. 31.

    The genetics of Caenorhabditis elegans. Genetics 77, 71–94 (1974)

  32. 32.

    & Specific interference by ingested dsRNA. Nature 395, 854 (1998)

  33. 33.

    et al. TOR deficiency in C. elegans causes developmental arrest and intestinal atrophy by inhibition of mRNA translation. Curr. Biol. 12, 1448–1461 (2002)

  34. 34.

    & Measuring Caenorhabditis elegans life span on solid media. J. Vis. Exp. 27, 1152 (2009)

  35. 35.

    & Regulation of organogenesis by the Caenorhabditis elegans FoxA protein PHA-4. Science 295, 821–825 (2002)

  36. 36.

    et al. C. elegans behavior of preference choice on bacterial food. Mol. Cells 28, 209–213 (2009)

  37. 37.

    , , & Target identification using drug affinity responsive target stability (DARTS). Curr. Protoc. Chem. Biol. 3, 163–180 (2011)

  38. 38.

    , & Identification of direct protein targets of small molecules. ACS Chem. Biol. 6, 34–46 (2011)

  39. 39.

    et al. Application of a proteolysis/mass spectrometry method for investigating the effects of inhibitors on hydroxylase structure. J. Med. Chem. 52, 2799–2805 (2009)

  40. 40.

    et al. High throughput microplate respiratory measurements using minimal quantities of isolated mitochondria. PLoS ONE 6, e21746 (2011)

  41. 41.

    Molecular Biology of the Cell 3rd edn (Garland, 1994)

  42. 42.

    et al. Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. Am. J. Physiol. Cell Physiol. 292, C125–C136 (2007)

  43. 43.

    et al. NCoR1 is a conserved physiological modulator of muscle mass and oxidative function. Cell 147, 827–839 (2011)

  44. 44.

    , , , & Coordinate regulation of lipid metabolism by novel nuclear receptor partnerships. PLoS Genet. 8, e1002645 (2012)

  45. 45.

    & The modular phosphorylation and activation of p70s6k. FEBS Lett. 410, 78–82 (1997)

  46. 46.

    , , , & RAFT1 phosphorylation of the translational regulators p70 S6 kinase and 4E-BP1. Proc. Natl Acad. Sci. USA 95, 1432–1437 (1998)

  47. 47.

    et al. Hierarchical phosphorylation of the translation inhibitor 4E-BP1. Genes Dev. 15, 2852–2864 (2001)

  48. 48.

    , , & Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307, 1098–1101 (2005)

  49. 49.

    , , & AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nature Cell Biol. 13, 132–141 (2011)

  50. 50.

    , & Dual roles of autophagy in the survival of Caenorhabditis elegans during starvation. Genes Dev. 21, 2161–2171 (2007)

  51. 51.

    et al. A role for autophagy in the extension of lifespan by dietary restriction in C. elegans. PLoS Genet. 4, e24 (2008)

  52. 52.

    , , & The autophagosomal protein LGG-2 acts synergistically with LGG-1 in dauer formation and longevity in C. elegans. Autophagy 6, 622–633 (2010)

  53. 53.

    , & NIH Image to ImageJ: 25 years of image analysis. Nature Methods 9, 671–675 (2012)

  54. 54.

    et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 19, 5720–5728 (2000)

  55. 55.

    et al. Cell-permeating α-ketoglutarate derivatives alleviate pseudohypoxia in succinate dehydrogenase-deficient cells. Mol. Cell. Biol. 27, 3282–3289 (2007)

  56. 56.

    et al. Glioma-derived mutations in IDH1 dominantly inhibit IDH1 catalytic activity and induce HIF-1α. Science 324, 261–265 (2009)

  57. 57.

    et al. Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Cancer Cell 19, 17–30 (2011)

  58. 58.

    et al. Disruption of wild-type IDH1 suppresses D-2-hydroxyglutarate production in IDH1-mutated gliomas. Cancer Res. 73, 496–501 (2013)

  59. 59.

    & Synthesis of the 1-monoester of 2-ketoalkanedioic acids, for example, octyl α-ketoglutarate. J. Org. Chem. 77, 11002–11005 (2012)

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We thank S. Lee, M. Hansen, B. Lemire, A. van der Bliek, S. Clarke, T. K. Blackwell, R. Johnson, J. E. Walker, A. G. W. Leslie, K. N. Houk, B. Martin, J. Lusis, J. Gober, Y. Wang and H. Sun for advice and discussions. J. Avruch for the let-363 RNAi vector; J. Powell-Coffman for strains and advice; and K. Yan for technical assistance. Worm strains were provided by the Caenorhabditis Genetics Center, which is funded by the National Institutes of Health (NIH) Office of Research Infrastructure Programs (P40 OD010440). We thank the NIH for traineeship support of R.M.C. (T32 GM007104), M.Y.P. (T32 GM007185), B.L. (T32 GM008496) and M.N. (T32 CA009120). X.F. is a recipient of the China Scholarship Council Scholarship. G.C.M. was supported by Ford Foundation and National Science Foundation Graduate Research Fellowships.

Author information

Author notes

    • Melody Y. Pai
    • , Laurent Vergnes
    •  & Heejun Hwang

    These authors contributed equally to this work.


  1. Molecular Biology Institute, University of California Los Angeles, Los Angeles, California 90095, USA

    • Randall M. Chin
    • , Melody Y. Pai
    • , Catherine F. Clarke
    • , Michael A. Teitell
    • , Karen Reue
    • , Michael E. Jung
    •  & Jing Huang
  2. Department of Molecular and Medical Pharmacology, University of California Los Angeles, Los Angeles, California 90095, USA

    • Xudong Fu
    • , Heejun Hwang
    • , Simon Diep
    • , Brett Lomenick
    • , Eileen Hu
    • , Gwanghyun Jung
    • , Austin Quach
    • , Abby S. Krall
    • , Meisheng Jiang
    • , Daniel Braas
    • , Heather R. Christofk
    •  & Jing Huang
  3. Department of Human Genetics, University of California Los Angeles, Los Angeles, California 90095, USA

    • Laurent Vergnes
    •  & Karen Reue
  4. Department of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, California 90095, USA

    • Gang Deng
    • , Catherine F. Clarke
    •  & Michael E. Jung
  5. Department of Biological Chemistry, University of California Los Angeles, Los Angeles, California 90095, USA

    • Vijaykumar S. Meli
    • , Gabriela C. Monsalve
    • , Feng Guo
    •  & Alison R. Frand
  6. Department of Surgery, University of California Los Angeles, Los Angeles, California 90095, USA

    • Stephen A. Whelan
    •  & Helena R. Chang
  7. Small Molecule Mass Spectrometry Facility, FAS Division of Science, Harvard University, Cambridge, Massachusetts 02138, USA

    • Jennifer X. Wang
    •  & Sunia A. Trauger
  8. Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA

    • Gregory M. Solis
    •  & Michael Petrascheck
  9. Pasarow Mass Spectrometry Laboratory, Department of Psychiatry and Biobehavioral Sciences and Semel Institute for Neuroscience and Human Behavior, University of California Los Angeles, Los Angeles, California 90095, USA

    • Farbod Fazlollahi
    •  & Kym F. Faull
  10. Department of Environmental Health Sciences, University of California Los Angeles, Los Angeles, California 90095, USA

    • Chitrada Kaweeteerawat
    •  & Hilary A. Godwin
  11. Department of Pathology and Laboratory Medicine, University of California Los Angeles, Los Angeles, California 90095, USA

    • Mahta Nili
    •  & Michael A. Teitell
  12. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA

    • Alan Saghatelian
  13. UCLA Metabolomics Center, University of California Los Angeles, Los Angeles, California 90095, USA

    • Daniel Braas
    •  & Heather R. Christofk


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Lifespan assays were performed by R.M.C., M.P. and E.H.; DARTS-mass spectrometry by S.D. and B.L.; DARTS-western blots by M.Y.P., H.H. and R.M.C.; mammalian cell experiments by X.F. and H.H.; mitochondrial respiration study design and analyses by L.V. and K.R.; enzyme kinetics and analyses by R.M.C. and J.H.; confocal microscopy by V.S.M., G.C.M. and A.R.F.; ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI/MS/MS) by J.X.W. and S.A.T.; compound syntheses by G.D. and M.E.J.; other analyses by H.H., X.F., M.Y.P., D.B., R.M.C., E.H., G.J., G.M.S., C.K. and A.Q. S.A.W., F.F., M.N., A.S.K., H.A.G., H.R. Chang, K.F.F., F.G., M.J., S.A.T., A.S., D.B., H.R. Christofk, C.F.C., M.A.T., M.E.J., L.V., K.R., A.R.F. and M.P. provided guidance, specialized reagents and expertise. J.H. conceived the study. R.M.C. and J.H. wrote the paper. R.M.C., X.F. and J.H. analysed data. All authors discussed the results, commented on the studies and contributed to aspects of preparing the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jing Huang.

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  1. 1.

    Example of a vehicle-treated day 16 adult animal

    The animal had lost all motility in the body and could only move its head slowly.

  2. 2.

    Example of a α-KG-treated day 16 adult animal

    The animal remained youthful and exhibited full body movements.

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