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RBX1-mediated ubiquitination of SESN2 promotes cell death upon prolonged mitochondrial damage in SH-SY5Y neuroblastoma cells

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Abstract

Sestrins are evolutionary conserved stress-inducible genes which regulate the axis of cell survival and cell death. Suppression of Sestrin 2 (SESN2) has been linked with increase in oxidative stress and cell death but mechanistic details related to regulation of SESN2 during mitochondrial damage remain unknown. Our study shows that prolonged CCCP-induced mitochondrial damage decreases SESN2 levels and viability of SH-SY5Y cells while overexpression of SESN2 significantly rescues the viability of cells. Further, we demonstrate that Ring box protein 1 (RBX1) is a novel interactive partner and E3 ligase for SESN2 which mediates its K-48-linked ubiquitination upon extensive mitochondrial damage. Downregulation of RBX1 causes stabilization in levels of SESN2. Notably, silencing of RBX1 expression substantially declines cell death and generation of mitochondrial ROS in response to prolonged mitochondrial damage. Taken together, we suggest that SESN2 is critical to protect cells against detrimental effect of mitochondrial damage and RBX1 is a negative regulator of SESN2 which hampers its stabilization.

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Abbreviations

SESN2:

Sestrin2

CCCP:

Carbonyl cyanide m-chlorophenylhydrazone

RBX1:

Ring box protein 1

AMPK:

AMP kinase

AD:

Alzheimer’s disease

PD:

Parkinson’s disease

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Acknowledgements

We acknowledge technical assistance from Mr. G.S. Neelaram. This work was supported by grants from the Department of Biotechnology, New Delhi, India (http://dbtindia.nic.in/index.asp), to the National Institute of Immunology (Grant No. BT/03/033/88); Centre for Molecular Medicine, New Delhi (Grant No. BT/PR/14549/MED/14/1291).

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Authors and Affiliations

  1. Cell Death and Differentiation Research Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Ashish Kumar & Chandrima Shaha

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  1. Ashish Kumar
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  2. Chandrima Shaha
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AK and CS formed the general framework of this study. AK and CS designed experiments. AK performed all experiments. AK and CS prepared the manuscript.

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Correspondence to Chandrima Shaha.

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Kumar, A., Shaha, C. RBX1-mediated ubiquitination of SESN2 promotes cell death upon prolonged mitochondrial damage in SH-SY5Y neuroblastoma cells. Mol Cell Biochem 446, 1–9 (2018). https://doi.org/10.1007/s11010-017-3267-7

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  • DOI: https://doi.org/10.1007/s11010-017-3267-7

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