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Lifespan extension induced by AMPK and calcineurin is mediated by CRTC-1 and CREB

Abstract

Activating AMPK or inactivating calcineurin slows ageing in Caenorhabditis elegans1,2 and both have been implicated as therapeutic targets for age-related pathology in mammals3,4,5. However, the direct targets that mediate their effects on longevity remain unclear. In mammals, CREB-regulated transcriptional coactivators (CRTCs)6 are a family of cofactors involved in diverse physiological processes including energy homeostasis7,8,9, cancer10 and endoplasmic reticulum stress11. Here we show that both AMPK and calcineurin modulate longevity exclusively through post-translational modification of CRTC-1, the sole C. elegans CRTC. We demonstrate that CRTC-1 is a direct AMPK target, and interacts with the CREB homologue-1 (CRH-1) transcription factor in vivo. The pro-longevity effects of activating AMPK or deactivating calcineurin decrease CRTC-1 and CRH-1 activity and induce transcriptional responses similar to those of CRH-1 null worms. Downregulation of crtc-1 increases lifespan in a crh-1-dependent manner and directly reducing crh-1 expression increases longevity, substantiating a role for CRTCs and CREB in ageing. Together, these findings indicate a novel role for CRTCs and CREB in determining lifespan downstream of AMPK and calcineurin, and illustrate the molecular mechanisms by which an evolutionarily conserved pathway responds to low energy to increase longevity.

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Figure 1: CRTC-1 regulates longevity.
Figure 2: CRTC-1 is a target of AAK-2 and TAX-6.
Figure 3: Calcineurin and AMPK regulate lifespan through phosphorylation of CRTC-1.
Figure 4: CREB activity regulates lifespan.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Data have been deposited at GEO under accession number GSE25513.

References

  1. Apfeld, J., O’Connor, G., McDonagh, T., DiStefano, P. S. & Curtis, R. The AMP-activated protein kinase AAK-2 links energy levels and insulin-like signals to lifespan in C. elegans . Genes Dev. 18, 3004–3009 (2004)

    Article  CAS  Google Scholar 

  2. Dong, M. Q. et al. Quantitative mass spectrometry identifies insulin signaling targets in C. elegans . Science 317, 660–663 (2007)

    Article  ADS  CAS  Google Scholar 

  3. Steinberg, G. R. & Kemp, B. E. AMPK in health and disease. Physiol. Rev. 89, 1025–1078 (2009)

    Article  CAS  Google Scholar 

  4. Supnet, C. & Bezprozvanny, I. Neuronal calcium signaling, mitochondrial dysfunction, and Alzheimer’s disease. J. Alzheimers Dis. 20 (suppl. 2). 487–498 (2010)

    Article  Google Scholar 

  5. Shackelford, D. B. & Shaw, R. J. The LKB1–AMPK pathway: metabolism and growth control in tumour suppression. Nature Rev. Cancer 9, 563–575 (2009)

    Article  CAS  Google Scholar 

  6. Conkright, M. D. et al. TORCs: transducers of regulated CREB activity. Mol. Cell 12, 413–423 (2003)

    Article  CAS  Google Scholar 

  7. Liu, Y. et al. A fasting inducible switch modulates gluconeogenesis via activator/coactivator exchange. Nature 456, 269–273 (2008)

    Article  ADS  CAS  Google Scholar 

  8. Screaton, R. A. et al. The CREB coactivator TORC2 functions as a calcium- and cAMP-sensitive coincidence detector. Cell 119, 61–74 (2004)

    Article  CAS  Google Scholar 

  9. Koo, S. H. et al. The CREB coactivator TORC2 is a key regulator of fasting glucose metabolism. Nature 437, 1109–1111 (2005)

    Article  ADS  CAS  Google Scholar 

  10. Komiya, T. et al. Enhanced activity of the CREB co-activator Crtc1 in LKB1 null lung cancer. Oncogene 29, 1672–1680 (2010)

    Article  CAS  Google Scholar 

  11. Wang, Y., Vera, L., Fischer, W. H. & Montminy, M. The CREB coactivator CRTC2 links hepatic ER stress and fasting gluconeogenesis. Nature 460, 534–537 (2009)

    Article  ADS  CAS  Google Scholar 

  12. Crute, B. E., Seefeld, K., Gamble, J., Kemp, B. E. & Witters, L. A. Functional domains of the α1 catalytic subunit of the AMP-activated protein kinase. J. Biol. Chem. 273, 35347–35354 (1998)

    Article  CAS  Google Scholar 

  13. Leffler, A. et al. The vanilloid receptor TRPV1 is activated and sensitized by local anesthetics in rodent sensory neurons. J. Clin. Invest. 118, 763–776 (2008)

    PubMed  PubMed Central  Google Scholar 

  14. Wang, W. & Shakes, D. C. Expression patterns and transcript processing of ftt-1 and ftt-2, two C. elegans 14-3-3 homologues. J. Mol. Biol. 268, 619–630 (1997)

    Article  CAS  Google Scholar 

  15. Kostelecky, B., Saurin, A. T., Purkiss, A., Parker, P. J. & McDonald, N. Q. Recognition of an intra-chain tandem 14-3-3 binding site within PKCε. EMBO Rep. 10, 983–989 (2009)

    Article  CAS  Google Scholar 

  16. Johnson, C. et al. Bioinformatic and experimental survey of 14-3-3-binding sites. Biochem. J. 427, 69–78 (2010)

    Article  CAS  Google Scholar 

  17. Kimura, Y. et al. A CaMK cascade activates CRE-mediated transcription in neurons of Caenorhabditis elegans . EMBO Rep. 3, 962–966 (2002)

    Article  CAS  Google Scholar 

  18. Mayr, B. & Montminy, M. Transcriptional regulation by the phosphorylation-dependent factor CREB. Nature Rev. Mol. Cell Biol. 2, 599–609 (2001)

    Article  CAS  Google Scholar 

  19. Xiao, X., Li, B. X., Mitton, B., Ikeda, A. & Sakamoto, K. M. Targeting CREB for cancer therapy: friend or foe. Curr. Cancer Drug Targets 10, 384–391 (2010)

    Article  CAS  Google Scholar 

  20. Urano, F. et al. A survival pathway for Caenorhabditis elegans with a blocked unfolded protein response. J. Cell Biol. 158, 639–646 (2002)

    Article  CAS  Google Scholar 

  21. Haskins, K. A., Russell, J. F., Gaddis, N., Dressman, H. K. & Aballay, A. Unfolded protein response genes regulated by CED-1 are required for Caenorhabditis elegans innate immunity. Dev. Cell 15, 87–97 (2008)

    Article  CAS  Google Scholar 

  22. Viswanathan, M., Kim, S. K., Berdichevsky, A. & Guarente, L. A role for SIR-2.1 regulation of ER stress response genes in determining C. elegans life span. Dev. Cell 9, 605–615 (2005)

    Article  CAS  Google Scholar 

  23. Greer, E. L. et al. An AMPK-FOXO pathway mediates longevity induced by a novel method of dietary restriction in C. elegans . Curr. Biol. 17, 1646–1656 (2007)

    Article  CAS  Google Scholar 

  24. Jansson, D. et al. Glucose controls CREB activity in islet cells via regulated phosphorylation of TORC2. Proc. Natl Acad. Sci. USA 105, 10161–10166 (2008)

    Article  ADS  Google Scholar 

  25. Erion, D. M. et al. Prevention of hepatic steatosis and hepatic insulin resistance by knockdown of cAMP response element-binding protein. Cell Metab. 10, 499–506 (2009)

    Article  CAS  Google Scholar 

  26. Shaw, R. J. et al. The kinase LKB1 mediates glucose homeostasis in liver and therapeutic effects of metformin. Science 310, 1642–1646 (2005)

    Article  ADS  CAS  Google Scholar 

  27. Qi, L. et al. Adipocyte CREB promotes insulin resistance in obesity. Cell Metab. 9, 277–286 (2009)

    Article  CAS  Google Scholar 

  28. Lerner, R. G., Depatie, C., Rutter, G. A., Screaton, R. A. & Balthasar, N. A role for the CREB co-activator CRTC2 in the hypothalamic mechanisms linking glucose sensing with gene regulation. EMBO Rep. 10, 1175–1181 (2009)

    Article  CAS  Google Scholar 

  29. Hope, I. A. C. elegans: A Practical Approach (ed. Hames. B. D.) (Oxford Univ. Press, 1999)

    Google Scholar 

  30. Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans . Nature 391, 806–811 (1998)

    Article  ADS  CAS  Google Scholar 

  31. Mair, W. A simple yet effective method to manipulate C. elegans in liquid. Worm Breed. Gaz. 18, 33 (2009)

    Google Scholar 

  32. Gentleman, R. C. et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80 (2004)

    Article  Google Scholar 

  33. Affymetrix. GeneChip Expression Analysishttp://www.coriell.org/images/pdf/expression_manual.pdf〉 (Affymetrix, 2004)

  34. Wu, Z., Irizarry, R. A., Gentleman, R., Martinez-Murillo, F. & Spencer, F. A model-based background adjustment for oligonucleotide expression arrays. J. Am. Stat. Assoc. 99, 909–917 (2004)

    Article  MathSciNet  Google Scholar 

  35. Smyth, G. K. Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Gen. Mol. Biol. 3, article 3. (2004)

    Article  MathSciNet  Google Scholar 

  36. Benjamini, Y. & Hochberg, Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Stat. Soc. B 57, 289–300 (1995)

    MathSciNet  MATH  Google Scholar 

  37. Kel, A. E. et al. MATCH: a tool for searching transcription factor binding sites in DNA sequences. Nucleic Acids Res. 31, 3576–3579 (2003)

    Article  CAS  Google Scholar 

  38. Matys, V. et al. TRANSFAC and its module TRANSCompel: transcriptional gene regulation in eukaryotes. Nucleic Acids Res. 34, D108–D110 (2006)

    Article  ADS  CAS  Google Scholar 

  39. Benbrook, D. M. & Jones, N. C. Different binding specificities and transactivation of variant CRE’s by CREB complexes. Nucleic Acids Res. 22, 1463–1469 (1994)

    Article  CAS  Google Scholar 

  40. Bucher, P. Weight matrix descriptions of four eukaryotic RNA polymerase II promoter elements derived from 502 unrelated promoter sequences. J. Mol. Biol. 212, 563–578 (1990)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

W.M. is funded by the George E. Hewitt Foundation for Medical Research, the American Federation for Aging Research and the Glenn Foundation for Medical Research. R.J.S. is funded by National Institutes of Health (NIH) R01 DK080425 and P01 CA120964. A.P.C.R. and G.M. are funded by NIH R01 HG004164, AG031097 and CA14195. A.D. is supported by NIH R01 DK070696 and AG027463. We thank the Caenorhabditis Genetics Center, the National Bioresource Project for the Nematode and Mark Alkema for providing worm strains. We are grateful to M. Raices and M. D’Angelo for critical analysis of the manuscript, DAPI images and the NUP-160::GFP construct. We also thank members of the A.D. laboratory and M. Hansen for comments on the manuscript and discussion and K. Butler for technical assistance in the early stages of this project.

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Authors

Contributions

W.M., I.M., M.M., R.J.S. and A.D. designed the experiments. W.M. and I.M. performed the experiments. A.P.C.R analysed the microarray data and performed the promoter analysis and W.M. analysed and performed statistical analysis on all other data. The manuscript was written by W.M. and edited by I.M., A.P.C.R., G.M., R.J.S. and A.D. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Reuben J. Shaw or Andrew Dillin.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-15 with legends and Supplementary Tables 3 and 4 (see Separate files for Supplementary Tables 1, 2 and 5). (PDF 14949 kb)

Supplementary Table 1

This table displays normalized expression measurements for all probesets. (PDF 24514 kb)

Supplementary Table 2

This table displays differentially expressed genes in each of the three mutants relative to wild-type. Each row correponds to a probeset, so a gene may appear in multiple rows. Also, reported are GO functional categories associated with each gene and CRE and TATA motifs identified upstream of each gene, the notation used follows the key: Hit Score | Palindromic | Hit Position | Hit Location in Genome | Distance to TSS | Conservation in other Caernohabditis. (XLS 2318 kb)

Supplementary Table 5

This table displays survival data and statistics for life span experiments. (XLS 16 kb)

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Mair, W., Morantte, I., Rodrigues, A. et al. Lifespan extension induced by AMPK and calcineurin is mediated by CRTC-1 and CREB. Nature 470, 404–408 (2011). https://doi.org/10.1038/nature09706

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