Abstract
The ubiquitous vacuolar H+-ATPase, a multisubunit proton pump, is essential for intraorganellar acidification. Disruption of its function leads to disturbances of organelle function and cell death. Here, we report that overexpression of the B2 subunit of the H+-ATPase inhibits apoptosis. This antiapoptotic effect is not mediated by an increase in H+-ATPase activity but through activation of the Ras-mitogen-activated protein kinase (MAPK)-signaling pathway that results in the serine phosphorylation of Bad at residues 112 and 155. Increased Bad phosphorylation reduces its translocation to mitochondria, limits the release of mitochondrial cytochrome c and apoptosis-inducing factor and increases the resistance of the B2 overexpressing cells to apoptosis. Screening experiments of kinase inhibitors, including inhibitors of cAMP-activated protein kinase, protein kinase C, protein kinase B, (MAPK/extracellular signal-regulated (ERK) kinase) MEK and Ste-MEK113, a cell permeable ERK activation inhibitor peptide, revealed that the B2 subunit of H+-ATPase acts upstream of MEK activation in the MEK/ERK pathway to ameliorate apoptosis.
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Abbreviations
- MAPK:
-
mitogen-activated protein kinase
- ERK:
-
extracellular-signal-regulated kinase
- MEK:
-
MAPK/ERK kinase
- RSK:
-
ribosomal S6 kinase
- PKA:
-
cAMP-activated protein kinase
- PKC:
-
protein kinase C
- AKT:
-
protein kinase B
- AIF:
-
apoptosis-inducing factor
- GFP:
-
green fluorescent protein
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Acknowledgements
This research is supported by grants obtained from National Institutes of Health (National Institute of Diabetes and Digestive and Kidney Diseases): JH Schwartz (DK-52898) and (DK 59529) and SC Borkan (DK-53387). We also appreciate the gift of rabbit antibody to the 31 kD subunit H+-ATPase from Dr Dennis Brown.
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Supplementary Information accompanies the paper on Cell Death and Differentiation website (http://www.nature.com/cdd)
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Li, G., Yang, Q., Krishnan, S. et al. A novel cellular survival factor – the B2 subunit of vacuolar H+-ATPase inhibits apoptosis. Cell Death Differ 13, 2109–2117 (2006). https://doi.org/10.1038/sj.cdd.4401970
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DOI: https://doi.org/10.1038/sj.cdd.4401970
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