Abstract
Sirtuins, a family of histone deacetylases, have a fiercely debated role in regulating lifespan. In contrast with recent observations, here we find that overexpression of sir-2.1, the ortholog of mammalian SirT1, does extend Caenorhabditis elegans lifespan. Sirtuins mandatorily convert NAD+ into nicotinamide (NAM). We here find that NAM and its metabolite, 1-methylnicotinamide (MNA), extend C. elegans lifespan, even in the absence of sir-2.1. We identify a previously unknown C. elegans nicotinamide-N-methyltransferase, encoded by a gene now named anmt-1, to generate MNA from NAM. Disruption and overexpression of anmt-1 have opposing effects on lifespan independent of sirtuins, with loss of anmt-1 fully inhibiting sir-2.1–mediated lifespan extension. MNA serves as a substrate for a newly identified aldehyde oxidase, GAD-3, to generate hydrogen peroxide, which acts as a mitohormetic reactive oxygen species signal to promote C. elegans longevity. Taken together, sirtuin-mediated lifespan extension depends on methylation of NAM, providing an unexpected mechanistic role for sirtuins beyond histone deacetylation.
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References
Nogueiras, R. et al. Sirtuins: physiological modulators of metabolism. Physiol. Rev. 92, 1479–1514 (2012).
Sinclair, D.A., Mills, K. & Guarente, L. Accelerated aging and nucleolar fragmentation in yeast sgs1 mutants. Science 277, 1313–1316 (1997).
Kaeberlein, M., McVey, M. & Guarente, L. The SIR2/3/4 complex and SIR2 alone promote longevity in Saccharomyces cerevisiae by two different mechanisms. Genes Dev. 13, 2570–2580 (1999).
Lin, S.J. et al. Calorie restriction extends Saccharomyces cerevisiae lifespan by increasing respiration. Nature 418, 344–348 (2002).
Anderson, R.M., Bitterman, K.J., Wood, J.G., Medvedik, O. & Sinclair, D.A. Nicotinamide and PNC1 govern lifespan extension by calorie restriction in Saccharomyces cerevisiae. Nature 423, 181–185 (2003).
Kaeberlein, M., Kirkland, K.T., Fields, S. & Kennedy, B.K. Sir2-independent life span extension by calorie restriction in yeast. PLoS Biol. 2, E296 (2004).
Schulz, T.J. et al. Glucose restriction extends Caenorhabditis elegans life span by inducing mitochondrial respiration and increasing oxidative stress. Cell Metab. 6, 280–293 (2007).
Tissenbaum, H.A. & Guarente, L. Increased dosage of a sir-2 gene extends lifespan in Caenorhabditis elegans. Nature 410, 227–230 (2001).
Viswanathan, M. & Guarente, L. Regulation of Caenorhabditis elegans lifespan by sir-2.1 transgenes. Nature 477, E1–E2 (2011).
Rizki,, G. et al. The evolutionarily conserved longevity determinants HCF-1 and SIR-2.1/SIRT1 collaborate to regulate DAF-16/FOXO. PLoS Genet 7, e1002235 (2011).
Rogina, B. & Helfand, S.L. Sir2 mediates longevity in the fly through a pathway related to calorie restriction. Proc. Natl. Acad. Sci. USA 101, 15998–16003 (2004).
Banerjee, K.K. et al. dSir2 in the adult fat body, but not in muscles, regulates life span in a diet-dependent manner. Cell Rep 2, 1485–1491 (2012).
Burnett, C. et al. Absence of effects of Sir2 overexpression on lifespan in C. elegans and Drosophila. Nature 477, 482–485 (2011).
Boily, G. et al. SirT1 regulates energy metabolism and response to caloric restriction in mice. PLoS One 3, e1759 (2008).
Kanfi, Y. et al. The sirtuin SIRT6 regulates lifespan in male mice. Nature 483, 218–221 (2012).
Gossmann, T.I. et al. NAD+ biosynthesis and salvage—a phylogenetic perspective. FEBS J. 279, 3355–3363 (2012).
Cantoni, G.L. Methylation of nicotinamide with soluble enzyme system from rat liver. J. Biol. Chem. 189, 203–216 (1951).
Fukushima, T. et al. Radical formation site of cerebral complex I and Parkinson′s disease. J. Neurosci. Res. 42, 385–390 (1995).
Ristow, M. & Zarse, K. How increased oxidative stress promotes longevity and metabolic health: The concept of mitochondrial hormesis (mitohormesis). Exp. Gerontol. 45, 410–418 (2010).
van der Horst, A., Schavemaker, J.M., Pellis-van Berkel, W. & Burgering, B.M. The Caenorhabditis elegans nicotinamidase PNC-1 enhances survival. Mech. Ageing Dev. 128, 346–349 (2007).
Hashimoto, T., Horikawa, M., Nomura, T. & Sakamoto, K. Nicotinamide adenine dinucleotide extends the lifespan of Caenorhabditis elegans mediated by sir-2.1 and daf-16. Biogerontology 11, 31–43 (2010).
Schmeisser, S. et al. Neuronal ROS signaling rather than AMPK/sirtuin-mediated energy sensing links dietary restriction to lifespan extension. Mol. Metab. 2, 92–102 (2013).
Kundu, T.K., Hille, R., Velayutham, M. & Zweier, J.L. Characterization of superoxide production from aldehyde oxidase: an important source of oxidants in biological tissues. Arch. Biochem. Biophys. 460, 113–121 (2007).
Sawyer, J.M. et al. Overcoming redundancy: an RNAi enhancer screen for morphogenesis genes in Caenorhabditis elegans. Genetics 188, 549–564 (2011).
Panoutsopoulos, G.I. & Beedham, C. Enzymatic oxidation of phthalazine with guinea pig liver aldehyde oxidase and liver slices: inhibition by isovanillin. Acta Biochim. Pol. 51, 943–951 (2004).
Crawford, D.l., Sutherland, J.B., Pometto, A.L., III & Miller, J.M. Production of an aromatic aldehyde oxidase by Streptomyces viridosporus. Arch. Microbiol. 131, 351–355 (1982).
Zarse, K. et al. Impaired insulin/IGF1-signaling extends life span by promoting mitochondrial l-proline catabolism to induce a transient ROS signal. Cell Metab. 15, 451–465 (2012).
Bishop, N.A. & Guarente, L. Two neurons mediate diet-restriction–induced longevity in C. elegans. Nature 447, 545–549 (2007).
Maeda, K. et al. Aldehyde oxidase 1 gene is regulated by Nrf2 pathway. Gene 505, 374–378 (2012).
Link, C.D. & Johnson, C.J. Reporter transgenes for study of oxidant stress in Caenorhabditis elegans. Methods Enzymol. 353, 497–505 (2002).
Van Raamsdonk, J.M. & Hekimi, S. Superoxide dismutase is dispensable for normal animal lifespan. Proc. Natl. Acad. Sci. USA 109, 5785–5790 (2012).
Dillin, A. et al. Rates of behavior and aging specified by mitochondrial function during development. Science 298, 2398–2401 (2002).
Yang, W. & Hekimi, S. Two modes of mitochondrial dysfunction lead independently to lifespan extension in Caenorhabditis elegans. Aging Cell 9, 433–447 (2010).
Ristow, M. et al. Antioxidants prevent health-promoting effects of physical exercise in humans. Proc. Natl. Acad. Sci. USA 106, 8665–8670 (2009).
Pan, Y., Schroeder, E.A., Ocampo, A., Barrientos, A. & Shadel, G.S. Regulation of yeast chronological life span by TORC1 via adaptive mitochondrial ROS signaling. Cell Metab. 13, 668–678 (2011).
Webster, B.R., Lu, Z., Sack, M.N. & Scott, I. The role of sirtuins in modulating redox stressors. Free Radic. Biol. Med. 52, 281–290 (2012).
Merksamer, P.I. et al. The sirtuins, oxidative stress and aging: an emerging link. Aging (Albany NY) 5, 144–150 (2013).
Brunet, A. et al. Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase. Science 303, 2011–2015 (2004).
Gebicki, J. & Chlopicki, S. Method for treating hypertriglyceridemia, dyslipidemia and hyperchlosterolemia with a 1-methylnicotinamide salt. US Patent 7,935,717 B2 10 (2011).
Nordestgaard, B.G., Benn, M., Schnohr, P. & Tybjaerg-Hansen, A. Triglycerides and risk of myocardial infarction, ischemic heart disease, and death in men and women. J. Am. Med. Assoc. 298, 299–308 (2007).
Brown, B.G. et al. Simvastatin and niacin, antioxidant vitamins, or the combination for the prevention of coronary disease. N. Engl. J. Med. 345, 1583–1592 (2001).
Canner, P.L. et al. Fifteen year mortality in Coronary Drug Project patients: long-term benefit with niacin. J. Am. Coll. Cardiol. 8, 1245–1255 (1986).
Warburton, D.E., Nicol, C.W. & Bredin, S.S. Health benefits of physical activity: the evidence. CMAJ 174, 801–809 (2006).
Chłopicki, S. et al. Single bout of endurance exercise increases NNMT activity in the liver and MNA concentration in plasma. Pharmacol. Rep. 64, 369–376 (2012).
Ulanovskaya, O.A., Zuhl, A.M. & Cravatt, B.F. NNMT promotes epigenetic remodeling in cancer by creating a metabolic methylation sink. Nat. Chem. Biol. 9, 300–306 (2013).
Parsons, R.B. et al. The expression of nicotinamide N-methyltransferase increases ATP synthesis and protects SH-SY5Y neuroblastoma cells against the toxicity of Complex I inhibitors. Biochem. J. 436, 145–155 (2011).
Mouchiroud, L. et al. The NAD+/Sirtuin pathway modulates longevity through activation of mitochondrial UPR and FOXO signaling. Cell 154, 430–441 (2013).
Segref, A. & Hoppe, T. Analysis of ubiquitin-dependent proteolysis in Caenorhabditis elegans. Methods Mol. Biol. 832, 531–544 (2012).
Janssen, A.J. et al. Spectrophotometric assay for complex I of the respiratory chain in tissue samples and cultured fibroblasts. Clin. Chem. 53, 729–734 (2007).
Musfeld, C., Biollaz, J., Belaz, N., Kesselring, U.W. & Decosterd, L.A. Validation of an HPLC method for the determination of urinary and plasma levels of N1-methylnicotinamide, an endogenous marker of renal cationic transport and plasma flow. J. Pharm. Biomed. Anal. 24, 391–404 (2001).
Trapnell, C., Pachter, L. & Salzberg, S.L. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25, 1105–1111 (2009).
Anders, S. & Huber, W. Differential expression analysis for sequence count data. Genome Biol. 11, R106 (2010).
Robinson, M.D., McCarthy, D.J. & Smyth, G.K. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139–140 (2010).
Priebe, S., Linde, J., Albrecht, D., Guthke, R. & Brakhage, A.A. FungiFun: a web-based application for functional categorization of fungal genes and proteins. Fungal Genet. Biol. 48, 353–358 (2011).
Wingender, E. The TRANSFAC project as an example of framework technology that supports the analysis of genomic regulation. Brief. Bioinform. 9, 326–332 (2008).
Blackwell, T.K., Bowerman, B., Priess, J.R. & Weintraub, H. Formation of a monomeric DNA binding domain by Skn-1 bZIP and homeodomain elements. Science 266, 621–628 (1994).
Acknowledgements
Most of the C. elegans strains used in this work were provided by the Caenorhabditis Genetics Center (University of Minnesota), which is funded by the US National Institutes of Health (NIH) Office of Research Infrastructure Programs (P40 OD010440). The strains LG389 and LG390 were a kind gift of L. Guarente and M. Viswanathan (both from Massachusetts Institute of Technology). The excellent technical assistance of I. Heinze, B. Laube, A. Müller, S. Richter and W. Scheiding as well as the excellent secretarial assistance of M. Schalowski are gratefully acknowledged. The RNA sequencing data contained in this manuscript were funded by the research program of the Jena Centre for Systems Biology of Ageing (JenAge) funded by the German Ministry for Education and Research (Bundesministerium für Bildung und Forschung; support code BMBF 0315581). D.A.S. is supported by grants from the NIH and National Institute on Aging, the United Mitochondrial Disease Foundation and the Glenn Medical Foundation.
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K.S. and J.M. designed, performed, and evaluated all of the experiments with the following exceptions: M.G. and M.P. performed next-generation sequencing analysis of mRNA, whereas sample provision, RNA extraction and quality control were done by K.S. Bioinformatical evaluation was done by S.P., R.G., I.H. and S. Schuster. Promoter analysis and gene classification was done by K.S. and J.M. A.S. and T.H. helped with strain constructions. S.W., D.K., A.P., M.B., S. Schmeisser, K.Z., N.L.P., Y.K., D.A.S. and H.Y.C. were involved in the study design and sample contribution and contributed several assays. The entire work was designed and supervised by M.R. The manuscript was written by K.S., J.M. and M.R. All of the authors discussed and commented on the manuscript.
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D.S. is a consultant and inventor on patents licensed to GlaxoSmithKline, PA, a company developing sirtuin-based medicines.
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Supplementary Results, Supplementary Figures 1–6 and Supplementary Tables 1 and 2. (PDF 1379 kb)
Supplementary Data Set 1
MNA-regulated genes (XLS 6906 kb)
Supplementary Data Set 2
NA-regulated genes (XLS 6815 kb)
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Schmeisser, K., Mansfeld, J., Kuhlow, D. et al. Role of sirtuins in lifespan regulation is linked to methylation of nicotinamide. Nat Chem Biol 9, 693–700 (2013). https://doi.org/10.1038/nchembio.1352
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DOI: https://doi.org/10.1038/nchembio.1352
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