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.

The bioenergetic and antioxidant status of neurons is controlled by continuous degradation of a key glycolytic enzyme by APC/C–Cdh1

Abstract

Neurons are known to have a lower glycolytic rate than astrocytes and when stressed they are unable to upregulate glycolysis1 because of low Pfkfb3 (6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase-3) activity2. This enzyme generates fructose-2,6-bisphosphate (F2,6P2)3, the most potent activator of 6-phosphofructo-1-kinase (Pfk1; ref. 4), a master regulator of glycolysis5. Here, we show that Pfkfb3 is absent from neurons in the brain cortex and that Pfkfb3 in neurons is constantly subject to proteasomal degradation by the action of the E3 ubiquitin ligase6, anaphase-promoting complex/cyclosome (APC/C)–Cdh1. By contrast, astrocytes have low APC/C–Cdh1 activity and therefore Pfkfb3 is present in these cells. Upregulation of Pfkfb3 by either inhibition of Cdh1 or overexpression of Pfkfb3 in neurons resulted in the activation of glycolysis. This, however, was accompanied by a marked decrease in the oxidation of glucose through the pentose phosphate pathway (a metabolic route involved in the regeneration of reduced glutathione7) resulting in oxidative stress and apoptotic death. Thus, by actively downregulating glycolysis by APC/C–Cdh1, neurons use glucose to maintain their antioxidant status at the expense of its utilization for bioenergetic purposes.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Pfkfb3 protein is degraded through the ubiquitin–proteasome pathway mediated by APC/C–Cdh1 in rat cortical neurons but not in astrocytes.
Figure 2: Cdh1 downregulates glycolysis and protects against apoptotic death through Pfkfb3 degradation in neurons.
Figure 3: Pfkfb3 expression in neurons triggers a decrease in glucose oxidation through the PPP, causing oxidative stress.
Figure 4: Expression of Pfkfb3 transiently protects neurons from loss of mitochondrial membrane potential (Δψm) and apoptotic death triggered by nitric oxide (NO).

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Almeida, A., Almeida, J., Bolaños, J. P. & Moncada, S. Different responses of astrocytes and neurons to nitric oxide: the role of glycolytically-generated ATP in astrocyte protection. Proc. Natl Acad. Sci. USA 98, 15294–15299 (2001).

    CAS  Article  Google Scholar 

  2. Almeida, A., Moncada, S. & Bolaños, J. P. Nitric oxide switches on glycolysis through the AMP protein kinase and 6-phosphofructo-2-kinase pathway. Nature Cell Biol. 6, 45–51 (2004).

    CAS  Article  Google Scholar 

  3. Hue, L. & Rider, M. H. Role of fructose 2, 6-bisphosphate in the control of glycolysis in mammalian tissues. Biochem. J. 245, 313–324 (1987).

    CAS  Article  Google Scholar 

  4. Van Schaftingen, E., Lederer, B., Bartrons, R. & Hers, H. G. A kinetic study of pyrophosphate: fructose-6-phosphate phosphotransferase from potato tubers. Application to a microassay of fructose 2, 6-bisphosphate. Eur. J. Biochem. 129, 191–195 (1982).

    CAS  Article  Google Scholar 

  5. Uyeda, K. Phosphofructokinase. Adv. Enzymol. Relat. Areas Mol. Biol. 48, 193–244 (1979).

    CAS  PubMed  Google Scholar 

  6. Sudakin, V. et al. The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis. Mol. Biol. Cell 6, 185–197 (1995).

    CAS  Article  Google Scholar 

  7. Kletzien, R. F., Harris, P. K. W. & Foellmi, L. A. Glucose-6-phosphate dehydrogenase: a housekeeping enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. FASEB J. 8, 174–181 (1994).

    CAS  Article  Google Scholar 

  8. Riera, L. et al. Regulation of ubiquitous 6-phosphofructo-2-kinase by the ubiquitin-proteasome proteolytic pathway during myogenic C2C12 cell differentiation. FEBS Lett. 550, 23–29 (2003).

    CAS  Article  Google Scholar 

  9. Pfleger, C. M. & Kirschner, M. W. The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1. Genes Dev. 14, 655–65 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Visintin, R., Prinz, S. & Amon, A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science 278, 460–463 (1997).

    CAS  Article  Google Scholar 

  11. Almeida, A., Bolanos, J. P. & Moreno, S. Cdh1/Hct1-APC is essential for the survival of postmitotic neurons. J. Neurosci. 25, 8115–8121 (2005).

    CAS  Article  Google Scholar 

  12. Cohen, S. S. Studies on the distribution of the oxidative pathway of glucose-6-phosphate utilization. Biol. Bull. 99, 369 (1950).

    CAS  Article  Google Scholar 

  13. Hothersall, J. S., Baquer, N. Z., Greenbaum, A. L. & McLean, P. Alternative pathways of glucose utilization in brain. Changes in the pattern of glucose utilization in brain during development and the effect of phenazine methosulphate on the integration of metabolic routes. Arch. Biochem. Biophys. 198, 478–492 (1979).

    CAS  Article  Google Scholar 

  14. Garcia Nogales, P., Almeida, A. & Bolaños, J. P. Peroxynitrite protects neurons against nitric oxide-mediated apoptosis. A key role for glucose-6-phosphate dehydrogenase activity in neuroprotection. J. Biol. Chem. 278, 864–874 (2003).

    CAS  Article  Google Scholar 

  15. Ben-Yoseph, O., Boxer, P. A. & Ross, B. D. Assessment of the role of the glutathione and pentose phosphate pathways in the protection of primary cerebrocortical cultures from oxidative stress. J. Neurochem. 66, 2329–2337 (1996).

    CAS  Article  Google Scholar 

  16. Vaughn, A. E. & Deshmukh, M. Glucose metabolism inhibits apoptosis in neurons and cancer cells by redox inactivation of cytochrome c. Nature Cell Biol. 10, 1477–1483 (2008).

    CAS  Article  Google Scholar 

  17. Tsacopoulos, M. & Magistretti, P. J. Metabolic coupling between glia and neurons. J. Neurosci. 16, 877–885 (1996).

    CAS  Article  Google Scholar 

  18. Pellerin, L. et al. Activity-dependent regulation of energy metabolism by astrocytes: an update. Glia 55, 1251–1262 (2007).

    Article  Google Scholar 

  19. Cidad, P., Almeida, A. & Bolaños, J. P. Inhibition of mitochondrial respiration by nitric oxide rapidly stimulates cytoprotective GLUT3-mediated glucose uptake through5′-AMP-activated protein kinase. Biochem. J. 384, 629–636 (2004).

    CAS  Article  Google Scholar 

  20. Brummelkamp, T. R., Bernards, R. & Agami, R. A system for stable expression of short interfering RNAs in mammalian cells. Science 296, 550–553 (2002).

    CAS  Article  Google Scholar 

  21. Reynolds, A. et al. Rational siRNA design for RNA interference. Nature Biotechnol. 22, 326–330 (2004).

    CAS  Article  Google Scholar 

  22. Ui-Tei, K. et al. Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res. 32, 936–948 (2004).

    CAS  Article  Google Scholar 

  23. Larrabee, M. G. Evaluation of the pentose phosphate pathway from 14CO2 data. Fallibility of a classic equation when applied to non-homogeneous tissues. Biochem. J. 272, 127–132 (1990).

    CAS  Article  Google Scholar 

  24. Lowry, O. H., Rosebrough, N. J., Lewis-Farr, A. & Randall, R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951).

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by Ministerio de Ciencia e Innovación (SAF2007-61492 and Consolider-Ingenio CSD2007-00020, Spain), Instituto de Salud Carlos-III (FIS06/0794 and Renevas), and Junta de Castilla y León (SA066A07 and Red de Terapia Celular y Medicina Regenerativa). We would like to thank H. Yamano for valuable help with the APC/C–Cdh1 activity assays, Carmela Gómez-Rodríguez for immunohistochemistry experiments, M. Resch for technical assistance and A. Higgs for critical evaluation of this paper.

Author information

Authors and Affiliations

Authors

Contributions

A.H.M., E.F. and C.M. performed the experiments; A.A. and J.P.B. analysed the data; A.A., S.M. and J.P.B. planned the experiments and wrote the paper.

Corresponding authors

Correspondence to Salvador Moncada or Juan P. Bolaños.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 4215 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Herrero-Mendez, A., Almeida, A., Fernández, E. et al. The bioenergetic and antioxidant status of neurons is controlled by continuous degradation of a key glycolytic enzyme by APC/C–Cdh1. Nat Cell Biol 11, 747–752 (2009). https://doi.org/10.1038/ncb1881

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1881

Further reading

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