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SIRT6 regulates TNF-α secretion through hydrolysis of long-chain fatty acyl lysine



The Sir2 family of enzymes or sirtuins are known as nicotinamide adenine dinucleotide (NAD)-dependent deacetylases1 and have been implicated in the regulation of transcription, genome stability, metabolism and lifespan2,3. However, four of the seven mammalian sirtuins have very weak deacetylase activity in vitro. Here we show that human SIRT6 efficiently removes long-chain fatty acyl groups, such as myristoyl, from lysine residues. The crystal structure of SIRT6 reveals a large hydrophobic pocket that can accommodate long-chain fatty acyl groups. We demonstrate further that SIRT6 promotes the secretion of tumour necrosis factor-α (TNF-α) by removing the fatty acyl modification on K19 and K20 of TNF-α. Protein lysine fatty acylation has been known to occur in mammalian cells, but the function and regulatory mechanisms of this modification were unknown. Our data indicate that protein lysine fatty acylation is a novel mechanism that regulates protein secretion. The discovery of SIRT6 as an enzyme that controls protein lysine fatty acylation provides new opportunities to investigate the physiological function of a protein post-translational modification that has been little studied until now.

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Figure 1: SIRT6 preferentially hydrolyses long-chain fatty acyl lysine in vitro.
Figure 2: Structure basis for SIRT6 activity with long-chain fatty acyl groups.
Figure 3: SIRT6 regulates TNF-α fatty acylation and secretion.

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Primary accessions

Protein Data Bank

Data deposits

The crystal structure of SIRT6 in complex with a H3K9 myristoyl peptide and ADP-ribose is deposited in the Protein Data Bank as accession number 3ZG6.


  1. Imai, S.-i., Armstrong, C. M., Kaeberlein, M. & Guarente, L. Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. Nature 403, 795–800 (2000)

    Article  ADS  CAS  Google Scholar 

  2. Sauve, A. A., Wolberger, C., Schramm, V. L. & Boeke, J. D. The biochemistry of sirtuins. Annu. Rev. Biochem. 75, 435–465 (2006)

    Article  CAS  Google Scholar 

  3. Michan, S. & Sinclair, D. Sirtuins in mammals: insights into their biological function. Biochem. J. 404, 1–13 (2007)

    Article  CAS  Google Scholar 

  4. Du, J. et al. Sirt5 is an NAD-dependent protein lysine demalonylase and desuccinylase. Science 334, 806–809 (2011)

    Article  ADS  CAS  Google Scholar 

  5. Zhu, A. Y. et al. Plasmodium falciparum Sir2A preferentially hydrolyzes medium and long chain fatty acyl lysine. ACS Chem. Biol. 7, 155–159 (2012)

    Article  CAS  Google Scholar 

  6. Mostoslavsky, R. et al. Genomic instability and aging-like phenotype in the absence of mammalian SIRT6. Cell 124, 315–329 (2006)

    Article  CAS  Google Scholar 

  7. Zhong, L. et al. The histone deacetylase Sirt6 regulates glucose homeostasis via Hif1α. Cell 140, 280–293 (2010)

    Article  CAS  Google Scholar 

  8. Xiao, C. et al. SIRT6 deficiency results in severe hypoglycemia by enhancing both basal and insulin-stimulated glucose uptake in mice. J. Biol. Chem. 285, 36776–36784 (2010)

    Article  CAS  Google Scholar 

  9. Michishita, E. et al. SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature 452, 492–496 (2008)

    Article  ADS  CAS  Google Scholar 

  10. Kanfi, Y. et al. The sirtuin SIRT6 regulates lifespan in male mice. Nature 483, 218–221 (2012)

    Article  ADS  CAS  Google Scholar 

  11. Yang, B., Zwaans, B. M. M., Eckersdorff, M. & Lombard, D. B. The sirtuin SIRT6 deacetylates H3 K56Ac in vivo to promote genomic stability. Cell Cycle 8, 2662–2663 (2009)

    Article  CAS  Google Scholar 

  12. Michishita, E. et al. Cell cycle-dependent deacetylation of telomeric histone H3 lysine K56 by human SIRT6. Cell Cycle 8, 2664–2666 (2009)

    Article  CAS  Google Scholar 

  13. Frye, R. A. Phylogenetic classification of prokaryotic and eukaryotic Sir2-like proteins. Biochem. Biophys. Res. Commun. 273, 793–798 (2000)

    Article  CAS  Google Scholar 

  14. Sebastián, C. et al. Deacetylase activity is required for STAT5-dependent GM-CSF functional activity in macrophages and differentiation to dendritic cells. J. Immunol. 180, 5898–5906 (2008)

    Article  Google Scholar 

  15. Hoff, K. G., Avalos, J. L., Sens, K. & Wolberger, C. Insights into the sirtuin mechanism from ternary complexes containing NAD+ and acetylated peptide. Structure 14, 1231–1240 (2006)

    Article  CAS  Google Scholar 

  16. Stevenson, F. T. et al. The 31-kDa precursor of interleukin 1 alpha is myristoylated on specific lysines within the 16-kDa N-terminal propiece. Proc. Natl Acad. Sci. USA 90, 7245–7249 (1993)

    Article  ADS  CAS  Google Scholar 

  17. Stevenson, F. T., Bursten, S. L., Locksley, R. M. & Lovett, D. H. Myristyl acylation of the tumor necrosis factor alpha precursor on specific lysine residues. J. Exp. Med. 176, 1053–1062 (1992)

    Article  CAS  Google Scholar 

  18. Locksley, R. M., Killeen, N. & Lenardo, M. J. The TNF and TNF receptor superfamilies: integrating mammalian biology. Cell 104, 487–501 (2001)

    Article  CAS  Google Scholar 

  19. Bruzzone, S. et al. Catastrophic NAD depletion in activated T lymphocytes through Nampt inhibition reduces demyelination and disability in EAE. PLoS ONE 4, e7897 (2009)

    Article  ADS  Google Scholar 

  20. Van Gool, F. et al. Intracellular NAD levels regulate tumor necrosis factor protein synthesis in a sirtuin-dependent manner. Nature Med. 15, 206–210 (2009)

    Article  CAS  Google Scholar 

  21. Charron, G. et al. Robust fluorescent detection of protein fatty-acylation with chemical reporters. J. Am. Chem. Soc. 131, 4967–4975 (2009)

    Article  CAS  Google Scholar 

  22. Wilson, J. P. et al. Proteomic analysis of fatty-acylated proteins in mammalian cells with chemical reporters reveals S-acylation of histone H3 variants. Mol. Cell. Proteomics 10, M110.001198 (2011)

    Article  Google Scholar 

  23. Martin, B. R. & Cravatt, B. F. Large-scale profiling of protein palmitoylation in mammalian cells. Nature Methods 6, 135–138 (2009)

    Article  CAS  Google Scholar 

  24. Schwer, B. et al. Neural sirtuin 6 (Sirt6) ablation attenuates somatic growth and causes obesity. Proc. Natl Acad. Sci. USA 107, 21790–21794 (2010)

    Article  ADS  CAS  Google Scholar 

  25. Walsh, C. T. Posttranslational Modification of Proteins: Expanding Nature’s Inventory (Roberts, 2005)

    Google Scholar 

  26. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997)

    Article  CAS  Google Scholar 

  27. Collaborative Computational Project, number 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50, 760–763 (1994)

  28. Pan, P. W. et al. Structure and biochemical functions of SIRT6. J. Biol. Chem. 286, 14575–14587 (2011)

    Article  CAS  Google Scholar 

  29. Stephens, S. B. et al. Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells. Methods Mol. Biol. 419, 197–214 (2008)

    Article  CAS  Google Scholar 

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This work was supported in part by NIH R01GM086703 (H.L.), R01GM093072 (R.M.), Hong Kong GRF766510 (Q.H.) and NIH R01GM087544 (H.C.H.). We thank C. Zhang for help with the cloning of SIRT6 WT and H133Y to generate lentiviral particles and the staff at the Shanghai Synchrotron Radiation Facility for assistance during the data collection.

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Authors and Affiliations



H.J. designed and carried out all biochemical experiments involving TNF-α and synthesized Rh-N3. S.K. synthesized acyl peptides and carried out all enzymology experiments of SIRT6. Y.W. carried out all crystallography experiments. G.C. and B.H. synthesized Alk14. G.C. and H.C.H. provided expertise on the labelling experiments using Alk14. C.S. and R.M. generated the MEF cells and bone marrow-derived macrophages from SIRT6 WT and KO mice. J.D., R.K. and E.G. contributed to the cloning, expression and purification of SIRT6. Q.H. directed the structural studies and wrote the structural part of the manuscript. H.L. directed the biochemical studies, coordinated the collaborations among different labs, and wrote the manuscript with help from H.J., S.K., Y.W., R.M., H.C.H. and Q.H.

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Correspondence to Quan Hao or Hening Lin.

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R.M. is on the scientific advisory board for Sirtris, a GSK company.

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Jiang, H., Khan, S., Wang, Y. et al. SIRT6 regulates TNF-α secretion through hydrolysis of long-chain fatty acyl lysine. Nature 496, 110–113 (2013).

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