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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.
Figure 2: Activation of PKB by CaM-KK in transfected COS-7 cells.
Figure 3: Phosphorylation of PKB and BAD as a result of CaM-KK activity in NG108 cells.
Figure 4: Regulation of apoptosis in NG108 cells.

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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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  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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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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