Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Regulation of the Caenorhabditis elegans longevity protein DAF-16 by insulin/IGF-1 and germline signaling

Nature Genetics volume 28, pages 139–145 (2001)Cite this article

Abstract

The lifespan of Caenorhabditis elegans is regulated by the insulin/insulin-like growth factor (IGF)-1 receptor homolog DAF-2, which signals through a conserved phosphatidylinositol 3-kinase (PI 3-kinase)/Akt pathway1,2,3,4,5,6,7. Mutants in this pathway remain youthful and active much longer than normal animals and can live more than twice as long. This lifespan extension requires DAF-16, a forkhead/winged-helix transcription factor8,9. DAF-16 is thought to be the main target of the DAF-2 pathway. Insulin/IGF-1 signaling is thought to lead to phosphorylation of DAF-16 by AKT activity, which in turn shortens lifespan. Here, we show that the DAF-2 pathway prevents DAF-16 accumulation in nuclei. Disrupting Akt-consensus phosphorylation sites in DAF-16 causes nuclear accumulation in wild-type animals, but, surprisingly, has little effect on lifespan. Thus the DAF-2 pathway must have additional outputs. Lifespan in C. elegans can be extended by perturbing sensory neurons or germ cells10,11. In both cases, lifespan extension requires DAF-16. We find that both sensory neurons and germline activity regulate DAF-16 accumulation in nuclei, but the nuclear localization patterns are different. Together these findings reveal unexpected complexity in the DAF-16-dependent pathways that regulate aging.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: DAF-16::GFP constructs.
Figure 2: Effect of mutant and wildtype DAF-16a:GFP proteins on lifespan.
Figure 3: Distribution of DAF-16a::GFP and DAF-16a::GFP/bKO proteins in L2 larvae and in dauers.
Figure 4: DAF-16aAM::GFP/bKO was localized in nuclei in both daf-16(mu86); daf-2(+) and daf-16(mu86); daf-2(e1370) backgrounds.
Figure 5: Localization of DAF-16a::GFP/bKO in cilium-structure sensory mutants and in germ-line ablated animals.
Figure 6: DAF-16a::GFP distribution in animals exposed to heat.

Similar content being viewed by others

References

  1. Kenyon, C., Chang, J., Gensch, E., Rudner, A. & Tabtiang, R. A C. elegans mutant that lives twice as long as wild type. Nature 366, 461–464 (1993).

    Article  CAS  Google Scholar 

  2. Morris, J.Z., Tissenbaum, H.A. & Ruvkun, G. A phosphatidylinositol-3-OH kinase family member regulating longevity and diapause in Caenorhabditis elegans. Nature 382, 536–539 (1996).

    Article  CAS  Google Scholar 

  3. Kimura, K.D., Tissenbaum, H.A., Liu, Y. & Ruvkun, G. daf-2, an insulin receptor-like gene that regulates longevity and diapause in Caenorhabditis elegans. Science 277, 942–946 (1997).

    Article  CAS  Google Scholar 

  4. Paradis, S. & Ruvkun, G. Caenorhabditis elegans Akt/PKB transduces insulin receptor-like signals from AGE-1 PI3 kinase to the DAF-16 transcription factor. Genes Dev. 12, 2488–2498 (1998).

    Article  CAS  Google Scholar 

  5. Paradis, S., Ailion, M., Toker, A., Thomas, J.H. & Ruvkun, G. A PDK1 homolog is necessary and sufficient to transduce AGE-1 PI3 kinase signals that regulate diapause in caenorhabditis elegans. Genes Dev. 13, 1438–1452 (1999).

    Article  CAS  Google Scholar 

  6. Ogg, S. & Ruvkun, G. The C. elegans PTEN homolog, DAF-18, acts in the insulin receptor-like metabolic signaling pathway. Mol. Cell 2, 887–893 (1998).

    Article  CAS  Google Scholar 

  7. Guarente, L. & Kenyon, C. Genetic pathways that regulate ageing in model organisms. Nature 408, 255–262 (2000).

    Article  CAS  Google Scholar 

  8. Ogg, S. et al. The fork head transcription factor DAF-16 transduces insulin-like metabolic and longevity signals in C. elegans. Nature 389, 994–999 (1997).

    Article  CAS  Google Scholar 

  9. Lin, K., Dorman, J.B., Rodan, A. & Kenyon, C. daf-16: An HNF-3-forkhead family member that can function to double the life-span of Caenorhabditis elegans. Science 278, 1319–1322 (1997).

    Article  CAS  Google Scholar 

  10. Apfeld, J. & Kenyon, C. Regulation of lifespan by sensory perception in caenorhabditis elegans. Nature 402, 804–809 (1999).

    Article  CAS  Google Scholar 

  11. Hsin, H. & Kenyon, C.J. Signals from the reproductive system regulate the lifespan of C. elegans. Nature 399, 362–366 (1999).

    Article  CAS  Google Scholar 

  12. Kenyon, C. A conserved regulatory system for aging. Cell 105, 165–168 (2001).

    Article  CAS  Google Scholar 

  13. Riddle, D.L. & Albert, P.S. in C. elegans II (eds. Riddle, D. L. et al.) 739–768 (Cold Spring Harbor Laboratory Press, NY, 1997).

    Google Scholar 

  14. Gil, E.B., Link, E.M., Liu, L.X., Johnson, C.D. & Lees, J.A. Regulation of the insulin-like developmental pathway of caenorhabditis elegans by a homolog of the PTEN tumor suppressor gene. Proc. Natl. Acad. Sci. USA 96, 2925–2930 (1999).

    Article  CAS  Google Scholar 

  15. Rouault, J.P. et al. Regulation of dauer larva development in caenorhabditis elegans by daf-18, a homologue of the tumour suppressor PTEN. Curr. Biol. 9, 329–332 (1999).

    Article  CAS  Google Scholar 

  16. Gottlieb, S. & Ruvkun, G. daf-2, daf-16 and daf-23: Genetically interacting genes controlling dauer formation in Caenorhabditis elegans. Genetics 137, 107–120 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Dorman, J.B., Albinder, B., Shroyer, T. & Kenyon, C. The age-1 and daf-2 genes function in a common pathway to control the lifespan of Caenorhabditis elegans. Genetics 141, 1399–1406 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Apfeld, J. & Kenyon, C. Cell nonautonomy of C. elegans daf-2 function in the regulation of diapause and life span. Cell 95, 199–210 (1998).

    Article  CAS  Google Scholar 

  19. Wolkow, C.A., Kimura, K.D., Lee, M.-S. & Ruvkun, G. Regulation of C. elegans life-span by insulinlike signaling in the nervous system. Science 290, 147–150 (2000).

    Article  CAS  Google Scholar 

  20. Vowels, J. & Thomas, J. Genetic analysis of chemosensory control of dauer formation in caenorhabditis elegans. Genetics 130, 105–123 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Brunet, A. et al. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96, 857–868 (1999).

    Article  CAS  Google Scholar 

  22. Kops, G.J. et al. Direct control of the Forkhead transcription factor AFX by protein kinase B. Nature 398, 630–634 (1999).

    Article  CAS  Google Scholar 

  23. Biggs III, W.H., Meisenhelder, J., Hunter, T., Cavenee, W.K. & Arden, K.C. Protein kinase B/Akt-mediated phosphorylation promotes nuclear exclusion of the winged helix transcription factor FKHR1. Proc. Natl. Acad. Sci. USA 96, 7421–7426 (1999).

    Article  Google Scholar 

  24. Nakae, J., Park, B.C. & Accili, D. Insulin stimulates phosphorylation of the forkhead transcription factor FKHR on serine 253 through a Wortmannin-sensitive pathway. J. Biol. Chem. 274, 15982–15985 (1999).

    Article  CAS  Google Scholar 

  25. Takaishi, H. et al. Regulation of nuclear translocation of Forkhead transcription factor AFX by protein kinase B. Proc. Natl. Acad. Sci. USA 96, 11836–11841 (1999).

    Article  CAS  Google Scholar 

  26. Guo, S. et al. Phosphorylation of serine 256 by protein kinase B disrupts transactivation by FKHR and mediates effects of insulin on insulin-like growth factor-binding protein-1 promoter activity through a conserved insulin response sequence. J. Biol. Chem. 274, 17184–17192 (1999).

    Article  CAS  Google Scholar 

  27. Tang, E.D., Nunez, G., Barr, F.G. & Guan, K.L. Negative regulation of the forkhead transcription factor FKHR by Akt. J. Biol. Chem. 274, 16741–16746 (1999).

    Article  CAS  Google Scholar 

  28. Malone, E.A., Inoue T. & Thomas, J. Genetic Analysis of the role of daf-28 and age-1 in regulating Caenorhabditis elegans dauer formation. Genetics 143, 1193–1205 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Gems, D. & Riddle, D.L. Defining wild-type life span in caenorhabditis elegans. J. Geront. 55A, B215–B219 (2000).

    Article  Google Scholar 

  30. Lithgow, G.J., White, T.M., Melov, S. & Johnson, T.E. Thermotolerance and extended life-span conferred by single-gene mutations and induced by thermal stress. Proc. Natl. Acad. Sci. USA 92, 7540–7544 (1995).

    Article  CAS  Google Scholar 

  31. Vallejo, A.N., Pogulis, R.J. & Pease, L.R. In vitro synthesis of novel genes: mutagenesis and recombination by PCR. PCR Methods and Application 4, S123–S130 (1994).

    Article  Google Scholar 

Download references

Acknowledgements

H.H. and N.L. contributed equally to this work. We thank B. Harmon, T. Yu, M. Singer, Q. Ch'ng and L. Yang for help with microscopy; B. Albinder for technical assistance; and all members of the Kenyon lab for discussions and comments on the manuscript. Some nematode strains were provided by the Caenorhabditis Genetics Center, which is supported by the National Center for Research Resources. This work was supported by a grant from the National Institutes of Health to C.K., who is the Herbert Boyer Professor of Biochemistry and Biophysics.

Author information

Author notes

  1. Kui Lin

    Present address: Department of Cell Biology and Technology, Genentech, Inc., South San Francisco, California, USA

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, USA

    Kui Lin, Honor Hsin, Natasha Libina & Cynthia Kenyon

Authors
  1. Kui Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Honor Hsin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natasha Libina
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cynthia Kenyon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cynthia Kenyon.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lin, K., Hsin, H., Libina, N. et al. Regulation of the Caenorhabditis elegans longevity protein DAF-16 by insulin/IGF-1 and germline signaling. Nat Genet 28, 139–145 (2001). https://doi.org/10.1038/88850

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/88850

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing