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BAD and glucokinase reside in a mitochondrial complex that integrates glycolysis and apoptosis

Abstract

Glycolysis and apoptosis are considered major but independent pathways that are critical for cell survival1,2,3,4. The activity of BAD, a pro-apoptotic BCL-2 family member, is regulated by phosphorylation in response to growth/survival factors5,6,7,8. Here we undertook a proteomic analysis to assess whether BAD might also participate in mitochondrial physiology. In liver mitochondria, BAD resides in a functional holoenzyme complex together with protein kinase A7 and protein phosphatase 1 (PP1) catalytic units9, Wiskott–Aldrich family member WAVE-1 as an A kinase anchoring protein10, and glucokinase (hexokinase IV)11. BAD is required to assemble the complex in that Bad-deficient hepatocytes lack this complex, resulting in diminished mitochondria-based glucokinase activity and blunted mitochondrial respiration in response to glucose. Glucose deprivation results in dephosphorylation of BAD, and BAD-dependent cell death. Moreover, the phosphorylation status of BAD helps regulate glucokinase activity. Mice deficient for BAD or bearing a non-phosphorylatable BAD(3SA) mutant12 display abnormal glucose homeostasis including profound defects in glucose tolerance. This combination of proteomics, genetics and physiology indicates an unanticipated role for BAD in integrating pathways of glucose metabolism and apoptosis.

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Figure 1: Characterization of mitochondrial complexes containing BAD.
Figure 2: Components of a mitochondrial BAD complex.
Figure 3: Aberrations in glucokinase activity, glucose homeostasis and glucose-withdrawal-induced cell death in Bad-/- mice.
Figure 4: Requirement of BAD phosphorylation in the regulation of mitochondrial glucokinase activity and glucose homeostasis.

References

  1. Raff, M. C. Social controls on cell survival and cell death. Nature 356, 397–400 (1992)

    ADS  CAS  Article  Google Scholar 

  2. Vander Heiden, M. G. et al. Growth factors can influence cell growth and survival through effects on glucose metabolism. Mol. Cell. Biol. 21, 5899–5912 (2001)

    CAS  Article  Google Scholar 

  3. Gottlob, K. et al. Inhibition of early apoptotic events by Akt/PKB is dependent on the first committed step of glycolysis and mitochondrial hexokinase. Genes Dev. 15, 1406–1418 (2001)

    CAS  Article  Google Scholar 

  4. Wang, X. The expanding role of mitochondria in apoptosis. Genes Dev. 15, 2922–2933 (2001)

    CAS  PubMed  Google Scholar 

  5. Zha, J., Harada, H., Yang, E., Jockel, J. & Korsmeyer, S. J. Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell 87, 619–628 (1996)

    CAS  Article  Google Scholar 

  6. Datta, S. R. et al. 14-3-3 proteins and survival kinases cooperate to inactivate BAD by BH3 domain phosphorylation. Mol. Cell 6, 41–51 (2000)

    CAS  Article  Google Scholar 

  7. Harada, H. et al. Phosphorylation and inactivation of BAD by mitochondria-anchored protein kinase A. Mol. Cell 3, 413–422 (1999)

    CAS  Article  Google Scholar 

  8. Harada, H., Andersen, J. S., Mann, M., Terada, N. & Korsmeyer, S. J. p70S6 kinase signals cell survival as well as growth, inactivating the pro-apoptotic molecule BAD. Proc. Natl Acad. Sci. USA 98, 9666–9670 (2001)

    ADS  CAS  Article  Google Scholar 

  9. Moorhead, G., MacKintosh, C., Morrice, N. & Cohen, P. Purification of the hepatic glycogen-associated form of protein phosphatase-1 by microcystin-Sepharose affinity chromatography. FEBS Lett. 362, 101–105 (1995)

    CAS  Article  Google Scholar 

  10. Westphal, R. S., Soderling, S. H., Alto, N. M., Langeberg, L. K. & Scott, J. D. Scar/WAVE-1, a Wiskott-Aldrich syndrome protein, assembles an actin-associated multi-kinase scaffold. EMBO J. 19, 4589–4600 (2000)

    CAS  Article  Google Scholar 

  11. Postic, C., Shiota, M. & Magnuson, M. A. Cell-specific roles of glucokinase in glucose homeostasis. Recent Prog. Horm. Res. 56, 195–217 (2001)

    CAS  Article  Google Scholar 

  12. Datta, S. R. et al. Survival factor-mediated BAD phosphorylation raises the mitochondrial threshold for apoptosis. Dev. Cell 3, 631–643 (2002)

    CAS  Article  Google Scholar 

  13. Wei, M. C. et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 292, 727–730 (2001)

    ADS  CAS  Article  Google Scholar 

  14. Cheng, E. H. et al. BCL-2, BCL-X(L) sequester BH3 domain-only molecules preventing BAX- and BAK-mediated mitochondrial apoptosis. Mol. Cell 8, 705–711 (2001)

    CAS  Article  Google Scholar 

  15. Hsu, Y. T. & Youle, R. J. Bax in murine thymus is a soluble monomeric protein that displays differential detergent-induced conformations. J. Biol. Chem. 273, 10777–10783 (1998)

    CAS  Article  Google Scholar 

  16. Krimmer, T. et al. Biogenesis of porin of the outer mitochondrial membrane involves an import pathway via receptors and the general import pore of the TOM complex. J. Cell Biol. 152, 289–300 (2001)

    CAS  Article  Google Scholar 

  17. Abdul, K. M. et al. Functional analysis of human metaxin in mitochondrial protein import in cultured cells and its relationship with the Tom complex. Biochem. Biophys. Res. Commun. 276, 1028–1034 (2000)

    CAS  Article  Google Scholar 

  18. Ayllon, V., Martinez, A. C., Garcia, A., Cayla, X. & Rebollo, A. Protein phosphatase 1α is a Ras-activated Bad phosphatase that regulates interleukin-2 deprivation-induced apoptosis. EMBO J. 19, 2237–2246 (2000)

    CAS  Article  Google Scholar 

  19. Westphal, R. S. et al. Regulation of NMDA receptors by an associated phosphatase-kinase signaling complex. Science 285, 93–96 (1999)

    CAS  Article  Google Scholar 

  20. Machesky, L. M. & Insall, R. H. Scar1 and the related Wiskott-Aldrich syndrome protein, WASP, regulate the actin cytoskeleton through the Arp2/3 complex. Curr. Biol. 8, 1347–1356 (1998)

    CAS  Article  Google Scholar 

  21. Postic, C. et al. Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase. J. Biol. Chem. 274, 305–315 (1999)

    CAS  Article  Google Scholar 

  22. Bali, D. et al. Animal model for maturity-onset diabetes of the young generated by disruption of the mouse glucokinase gene. J. Biol. Chem. 270, 21464–21467 (1995)

    CAS  Article  Google Scholar 

  23. Cho, H. et al. Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta). Science 292, 1728–1731 (2001)

    ADS  CAS  Article  Google Scholar 

  24. Plas, D. R., Talapatra, S., Edinger, A. L., Rathmell, J. C. & Thompson, C. B. Akt and Bcl-xL promote growth factor-independent survival through distinct effects on mitochondrial physiology. J. Biol. Chem. 276, 12041–12048 (2001)

    CAS  Article  Google Scholar 

  25. Garland, J. M. & Halestrap, A. Energy metabolism during apoptosis. Bcl-2 promotes survival in hematopoietic cells induced to apoptose by growth factor withdrawal by stabilizing a form of metabolic arrest. J. Biol. Chem. 272, 4680–4688 (1997)

    CAS  Article  Google Scholar 

  26. Murata, T. et al. Co-localization of glucokinase with actin filaments. FEBS Lett. 406, 109–113 (1997)

    CAS  Article  Google Scholar 

  27. Dekker, P. J. et al. The Tim core complex defines the number of mitochondrial translocation contact sites and can hold arrested preproteins in the absence of matrix Hsp70-Tim44. EMBO J. 16, 5408–5419 (1997)

    CAS  Article  Google Scholar 

  28. Licklider, L. J., Thoreen, C. C., Peng, J. & Gygi, S. P. Automation of nanoscale microcapillary liquid chromatography-tandem mass spectrometry with a vented column. Anal. Chem. 74, 3076–3083 (2002)

    CAS  Article  Google Scholar 

  29. Niswender, K. D. et al. Cell-specific expression and regulation of a glucokinase gene locus transgene. J. Biol. Chem. 272, 22564–22569 (1997)

    CAS  Article  Google Scholar 

  30. Zhang, C. Y. et al. Uncoupling protein-2 negatively regulates insulin secretion and is a major link between obesity, beta cell dysfunction, and type 2 diabetes. Cell 105, 745–755 (2001)

    CAS  Article  Google Scholar 

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Acknowledgements

We thank M. Ryan, G. Shore, J. Scott, M. Magnuson and B. Spiegelman for reagents; M. Ryan and J. Opferman for technical advice; B. Kahn and O. Peroni for discussion; S. Wade, J. Fisher and J. Sturgill for animal care; U. Maduekwe for technical assistance; and E. Smith for manuscript preparation. N.N.D. is a recipient of the Cancer Research Fund of Damon Runyon Foundation fellowship. This work is supported in part by a NIH grant.

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Correspondence to Stanley J. Korsmeyer.

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Danial, N., Gramm, C., Scorrano, L. et al. BAD and glucokinase reside in a mitochondrial complex that integrates glycolysis and apoptosis. Nature 424, 952–956 (2003). https://doi.org/10.1038/nature01825

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