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Nucleophosmin is a novel Bax chaperone that regulates apoptotic cell death

Oncogene volume 26, pages 2554–2562 (2007)Cite this article

Abstract

The proapoptotic B-cell lymphoma-2 family protein Bax is a key regulatory point in the intrinsic apoptotic pathway. However, the factors controlling the process of Bax activation and translocation to mitochondria have yet to be fully identified and characterized. We performed affinity chromatography using peptides corresponding to the mitochondrial-targeting region of Bax, which is normally sequestered within the inactive structure. The molecular chaperone nucleophosmin was identified as a novel Bax-binding protein by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Reciprocal co-immunoprecipitation and proximity assays confirmed the Bax-nucleophosmin protein–protein interaction and verified that nucleophosmin only bound to activated conformationally altered Bax. Confocal microscopy in a cell-based apoptosis model, demonstrated that nucleophosmin translocation from nucleolus to cytosol preceded Bax movement. Specific knockdown of nucleophosmin expression using RNAi attenuated apoptosis as measured by mitochondrial cytochrome c release and activation of the caspase cascade. In a mouse model of ischaemic stroke, subcellular fractionation studies verified that nucleophosmin translocation occurred within 3 h, at a time before Bax translocation but after Bax conformational changes have occurred. Thus, we have elucidated a novel molecular mechanism whereby Bax becomes activated and translocates to the mitochondria to orchestrate mitochondrial dysfunction and apoptotic cell death, which opens new avenues for therapeutic intervention.

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Acknowledgements

This work was funded by a research grant from Astellas Pharma Inc., Tokyo, Japan.

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  1. A L McGregor

    Present address: Department of Pharmacology, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand

Authors and Affiliations

  1. Astellas CNS Research in Edinburgh, The University of Edinburgh, Edinburgh, UK

    L E Kerr, J-LA Birse-Archbold, D M Short, A L McGregor, I Heron, D C MacDonald, J Thompson, G J Carlson, J S Kelly, J McCulloch & J Sharkey

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Correspondence to L E Kerr.

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Kerr, L., Birse-Archbold, JL., Short, D. et al. Nucleophosmin is a novel Bax chaperone that regulates apoptotic cell death. Oncogene 26, 2554–2562 (2007). https://doi.org/10.1038/sj.onc.1210044

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