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Reactive oxygen species induced by proteasome inhibition in neuronal cells mediate mitochondrial dysfunction and a caspase-independent cell death

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Abstract

While increasing evidence shows that proteasome inhibition triggers oxidative damage, mitochondrial dysfunction and death in neuronal cells, the regulatory relationship among these events is unclear. Using mouse neuronal cells we show that the cytotoxicity induced by mild (0.25 μM) and potent (5.0 μM) doses of the proteasome inhibitor, N-Benzyloxycarbonyl-Ile-Glu (O-t-butyl)-Ala-leucinal, (PSI) involved a dose-dependent increase in caspase activation, overproduction of reactive oxygen species (ROS) and a mitochondrial dysfunction manifested by the translocation of the proapoptotic protein, Bax, from the cytoplasm to the mitochondria, membrane depolarization and the release of cytochrome c and the apoptosis inducing factor (AIF) from mitochondria to the cytoplasm and nucleus, respectively. Whereas caspase or Bax inhibition failed to prevent mitochondrial membrane depolarization and neuronal cell death, pretreatments with the antioxidant N-acetyl-l-cysteine (NAC) or overexpression of the antiapoptotic protein Bcl-xL abrogated these events in cells exposed to mild levels of PSI. These findings implicated ROS as a mediator of PSI-induced cytotoxicity. However, depletions in glutathione and Bcl-xL with potent proteasome inhibition exacerbated this response whereupon survival required the cooperative protection of NAC with Bcl-xL overexpression. Collectively, ROS induced by proteasome inhibition mediates a mitochondrial dysfunction in neuronal cells that culminates in death through caspase- and Bax-independent mechanisms.

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Abbreviations

PSI:

N-Benzyloxycarbonyl-Ile-Glu (O-t-butyl)-Ala-leucinal

AIF:

Apoptosis inducing factor

ROS:

Reactive oxygen species

NAC:

N-acetyl-cysteine

GSH:

Glutathione

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Acknowledgements

This project was supported by The National Institutes of Health Grant (NIGMS SCORE to P.R.) and Grant Number RR03037 from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NCRR or NIH.

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  1. Department of Biological Sciences, Hunter College of The City University of New York, 695 Park Ave, New York, NY, 10021, USA

    Luena Papa, Evan Gomes & Patricia Rockwell

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  1. Luena Papa
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  2. Evan Gomes
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Correspondence to Patricia Rockwell.

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Papa, L., Gomes, E. & Rockwell, P. Reactive oxygen species induced by proteasome inhibition in neuronal cells mediate mitochondrial dysfunction and a caspase-independent cell death. Apoptosis 12, 1389–1405 (2007). https://doi.org/10.1007/s10495-007-0069-5

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