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Calcium promotes cell survival through CaM-K kinase activation of the protein-kinase-B pathway

Nature volume 396, pages 584–587 (1998)Cite this article

Abstract

The protection against apoptosis provided by growth factors in several cell lines is due to stimulation of the phosphatidylinositol-3-OH kinase (PI(3)K) pathway, which results in activation of protein kinase B1,2 (PKB; also known as c-Akt and Rac) and phosphorylation and sequestration to protein 14-3-3 of the pro-apoptotic Bcl-2-family member BAD3,4,5,6,7. A modest increase in intracellular Ca2+ concentration also promotes survival of some cultured neurons8,9 through a pathway that requires calmodulin but is independent of PI(3)K and the MAP kinases10,11. Here we report that Ca2+/calmodulin-dependent protein kinase kinase (CaM-KK) activates PKB directly, resulting in phosphorylation of BAD on serine residue 136 and the interaction of BAD with protein 14-3-3. Serum withdrawal induced a three- to fourfold increase in cell death of NG108 neuroblastoma cells, and this apoptosis was largely blocked by increasing the intracellular Ca2+ concentration with NMDA (N-methyl-D-aspartate) or KCl or by transfection with constitutively active CaM-KK. The effect of NMDA on cell survival was blocked by transfection with dominant-negative forms of CaM-KK or PKB. These results identify a Ca2+-triggered signalling cascade in which CaM-KK activates PKB, which in turn phosphorylates BAD and protects cells from apoptosis.

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Figure 1: Phosphorylation and activation of recombinant PKB by CaM-KK in vitro.
The alternative text for this image may have been generated using AI.
Figure 2: Activation of PKB by CaM-KK in transfected COS-7 cells.
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Figure 3: Phosphorylation of PKB and BAD as a result of CaM-KK activity in NG108 cells.
The alternative text for this image may have been generated using AI.
Figure 4: Regulation of apoptosis in NG108 cells.
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References

  1. Franke, T. F. et al. The protein kinase encoded by Akt proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase. Cell 81, 727–736 (1995).

    Article  CAS  Google Scholar 

  2. Burgering, B. M. T. & Coffer, P. J. Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature 376, 599–602 (1995).

    Article  ADS  CAS  Google Scholar 

  3. Datta, S. R. et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91, 231–241 (1997).

    Article  CAS  Google Scholar 

  4. Peso, L., Gonzalez-Garcia, M., Page, C., Herrera, R. & Nunez, G. Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. Science 278, 687–689 (1997).

    Article  ADS  Google Scholar 

  5. Hemmings, B. A. Akt signaling: linking membrane events to life and death decisions. Science 275, 628–630 (1997).

    Article  CAS  Google Scholar 

  6. Marte, B. M. & Downward, J. PKB/Akt: connecting phosphoinositide 3-kinase to cell survival and beyond. Trends Biochem. Sci. 22, 355–358 (1997).

    Article  CAS  Google Scholar 

  7. Dudek, H. et al. Regulation of neuronal survival by the serine-threonine protein kinase Akt. Science 275, 661–664 (1997).

    Article  CAS  Google Scholar 

  8. Gallo, V., Kingsbury, A., Balazs, R. & Jorgensen, O. S. The role of depolarization in the survival and differentiation of cerebellar granule cells in culture. J. Neurosci. 7, 2203–2213 (1987).

    Article  CAS  Google Scholar 

  9. Koike, T., Martin, D. P. & Johnson, E. M. Role of calcium channels in the ability of membrane-depolarization to prevent neuronal death induced by trophic factor deprivation: evidence that levels of internal Ca2+ determine nerve growth factor dependence of sympathetic ganglion cells. Proc. Natl Acad. Sci. USA 86, 6421–6425 (1989).

    Article  ADS  CAS  Google Scholar 

  10. Hack, N., Hidaka, H., Wakefield, M. J. & Balazs, R. Promotion of granule cell survival by high K+ or excitatory amino acid treatment and Ca2+/calmodulin-dependent protein kinase activity. Neuroscience 57, 90–20 (1993).

    Article  Google Scholar 

  11. Soler, R. M. et al. Calmodulin is involved in membrane depolarization-mediated survival of motoneurons by phosphatidylinositol-3 kinase- and MAPK-independent pathways. J. Neurosci. 18, 1230–1239 (1998).

    Article  CAS  Google Scholar 

  12. Alessi, D. R. et al. Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activated protein kinase Bα. Curr. Biol. 7, 261–269 (1997).

    Article  CAS  Google Scholar 

  13. Stokoe, D. et al. Dual role of phosphatidylinositol-3,4,5-trisphosphate in the activation of protein kinase B. Science 277, 567–570 (1997).

    Article  CAS  Google Scholar 

  14. Stephens, L. et al. Protein kinase B kinases that mediate phosphatidylinositol 3,4,5-trisphosphate-dependent activation of protein kinase B. Science 279, 710 (1998).

    Article  ADS  CAS  Google Scholar 

  15. Alessi, D. R. et al. Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO J. 15, 6541–6551 (1996).

    Article  CAS  Google Scholar 

  16. Haribabu, B. et al. Human calcium-calmodulin dependent protein kinase I: cDNA cloning, domain structure and activation by phosphorylation at threonine-177 by calcium-calmodulin dependent protein kinase I kinase. EMBO J. 14, 3679–3686 (1995).

    Article  CAS  Google Scholar 

  17. Sebert, M. A. et al. Phosphorylation and activation of Ca(2+)-calmodulin-dependent protein kinase IV by Ca(2+)-calmodulin-dependent protein kinase Ia kinase. Phosphorylation of threonine 196 is essential for activation. J. Biol. Chem. 270, 17616–17621 (1995).

    Article  Google Scholar 

  18. Tokumitsu, H., Enslen, H. & Soderling, T. R. Characterization of a CaM-kinase cascade: molecular cloning and characterization of CaM-kinase. J. Biol. Chem. 270, 19320–19324 (1995).

    Article  CAS  Google Scholar 

  19. Wayman, G. A., Tokumitsu, H. & Soderling, T. R. Inhibitory cross-talk by cAMP-kinase on the calmodulin-dependent protein kinase cascade. J. Biol. Chem. 272, 16073–16076 (1997).

    Article  CAS  Google Scholar 

  20. Conus, N. M., Hemmings, B. A. & Pearson, R. B. Differential regulation by calcium reveals distinct signaling requirements for the activation of akt and p70. J. Biol. Chem. 273, 4776–4782 (1998).

    Article  CAS  Google Scholar 

  21. Enslen, H., Tokumitsu, H., Stork, P. J. S., Davis, R. J. & Soderling, T. R. Regulation of MAP kinases by a calcium/calmodulin-dependent protein kinase cascade. Proc. Natl Acad. Sci. USA 93, 10803–10808 (1996).

    Article  ADS  CAS  Google Scholar 

  22. 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-XL. Cell 87, 619–628 (1996).

    Article  CAS  Google Scholar 

  23. Hsu, S. Y., Kaipia, A., Zhu, L. & Hsueh, A. J. Interference of BAD (Bcl-xL/Bcl-2-associated death promoter)-induced apoptosis in mammalian cells by 14-3-3 isoforms and P11. Mol. Endocrinol. 11, 1858–1867 (1997).

    CAS  PubMed  Google Scholar 

  24. Furbre, S., Oppenheim, R. W. & Prevette, D. Naturally-occuring neuron death in the ciliary ganglion of the chick embryo following removal of pregangalionic input: evidence for the role of afferents in ganglion cell survival. J. Neurosci. 7, 1816–1832 (1987).

    Article  Google Scholar 

  25. Coffer, P. J. & Woodgett, J. R. Molecular cloning and characterization of an novel putative protein-serine kinase related to the cAMP-dependent and protein kinase C families. Eur. J. Biochem. 201, 475–481 (1991).

    Article  CAS  Google Scholar 

  26. Jones, P. F., Jakubowicz, T., Pitossi, F. J., Maurer, F. & Hemmings, B. A. Molecular cloning and identification of a serine/threonine protein kinase of the second-messenger subfamily. Proc. Natl Acad. Sci. USA 88, 4171–4175 (1991).

    Article  ADS  CAS  Google Scholar 

  27. Tokumitsu, H. & Soderling, T. R. Requirements for calcium and calmodulin in the calmodulin-kinase activation cascade. J. Biol. Chem. 271, 5617–5622 (1996).

    Article  CAS  Google Scholar 

  28. Park, J. K. & Soderling, T. R. Activation of Ca2+/calmodulin-dependent protein kinase IV by CaM-kinase kinase in Jurkat T lymphocytes. J. Biol. Chem. 270, 30464–30469 (1995).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank B. Chang for help in constructing dominant-negative PKB and P. Stork for discussions during the course of this work. This study was supported by the NIH.

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Authors and Affiliations

  1. Vollum Institute, Oregon Health Sciences University, Portland, 97201, Oregon, USA

    Shigetoshi Yano, Hiroshi Tokumitsu & Thomas R. Soderling

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  1. Shigetoshi Yano
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  2. Hiroshi Tokumitsu
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  3. Thomas R. Soderling
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Correspondence to Thomas R. Soderling.

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Yano, S., Tokumitsu, H. & Soderling, T. Calcium promotes cell survival through CaM-K kinase activation of the protein-kinase-B pathway. Nature 396, 584–587 (1998). https://doi.org/10.1038/25147

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