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Non-canonical function of Bax in stress-induced nuclear protein redistribution

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Abstract

Bax and Bak (Bax/Bak) are essential pro-apoptotic proteins of the Bcl-2 family that trigger mitochondrial outer membrane permeabilization (MOMP) in a Bcl-2/Bcl-xL-inhibitable manner. We recently discovered a new stress-related function for Bax/Bak—regulation of nuclear protein redistribution (NPR) from the nucleus to cytoplasm. This effect was independent of Bax/Bak N-terminus exposure and not inhibited by Bcl-xL over-expression. Here, we studied the molecular mechanism governing this novel non-canonical response. Wild-type (WT) and mutant versions of Bax were re-expressed in Bax/Bak double-knockout mouse embryonic fibroblasts and their ability to promote NPR, apoptotic events, and changes in lamin A mobility was examined. Our results show that, in this system, Bax expression was sufficient to restore NPR such as in WT cells undergoing apoptosis. This activity of Bax was uncoupled from cytochrome c release from the mitochondria (indicative of MOMP) and required its membrane localization, α helices 5/6, and the Bcl-2 homology 3 (BH3) domain. Moreover, enrichment of Bax in the nuclear envelope by the so-called Klarsicht/ANC-1/Syne-1 homology domain effectively triggered NPR as in WT Bax, but without inducing MOMP or cell death. Bax-induced NPR was associated with impairment in lamin A mobility, implying a connection between these two nuclear envelope-associated events. Overall, the results indicate a new MOMP-independent, stress-induced Bax function on the nuclear envelope.

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Abbreviations

Bax/Bak:

Bax and Bak

DKO:

Double-knockout

DMEM:

Dulbecco’s modified Eagle’s medium

FRAP:

Fluorescence recovery after photobleaching

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

GFP:

Green fluorescent protein

HA:

Hemagglutinin

KASH:

Klarsicht/ANC-1/Syne-1 homology

MEFs:

Mouse embryonic fibroblasts

MOMP:

Mitochondrial outer membrane permeabilization

NE:

Nuclear envelope

NPR:

Nuclear protein redistribution

NPM:

Nucleophosmin

Q-VD-OPH:

Quinoline-val-asp(OMe)-CH2-OPH

RFP:

Red fluorescent protein

WT:

Wild-type

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Acknowledgments

We thank Prof. Andreas Strasser, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia, for providing the Bax/Bak DKO MEFs, Prof. Xu Luo, Eppley Institute for Cancer Research, University of Nebraska Medical Center, Omaha, NE, USA, for providing the GFP-Bax 63–65A and GFP-Bax 92–95A plasmids, Prof. Richard J. Youle, Surgical Neurology Branch, NINDS, National Institutes of Health, Bethesda, MD, USA, for providing the Bax S184V plasmid, and Prof. Howard J. Worman, Columbia University, NY, USA, for providing the lamin A-RFP and GFP-mini-nesprin 2G plasmids. This work was supported by the German-Israeli Foundation (to R. Stein and C. Borner), the Spemann Graduate School of Biology and Medicine (SGBM) (GSC-4) and the Excellence Cluster BIOSS (EXC-294) funded by the DFG (to C. Borner).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel Aviv University, 69978, Ramat Aviv, Israel

    Liora Lindenboim & Reuven Stein

  2. Department of Biology, Bioimaging Center, University of Konstanz, 78457, Constance, Germany

    Elisa Ferrando-May

  3. Institute of Molecular Medicine and Cell Research, Albert Ludwigs University Freiburg, Stefan Meier Strasse 17, 79104, Freiburg, Germany

    Christoph Borner

  4. Spemann Graduate School of Biology and Medicine (SGBM), Albert Ludwigs University Freiburg, Albertstrasse 19a, 79104, Freiburg, Germany

    Christoph Borner

  5. Excellence Cluster, Centre for Biological Signaling Studies (BIOSS), Albert Ludwigs University Freiburg, Hebelstrasse 25, 79104, Freiburg, Germany

    Christoph Borner

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  1. Liora Lindenboim
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  2. Elisa Ferrando-May
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  4. Reuven Stein
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Correspondence to Reuven Stein.

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Lindenboim, L., Ferrando-May, E., Borner, C. et al. Non-canonical function of Bax in stress-induced nuclear protein redistribution. Cell. Mol. Life Sci. 70, 3013–3027 (2013). https://doi.org/10.1007/s00018-013-1306-4

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  • DOI: https://doi.org/10.1007/s00018-013-1306-4

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