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Inhibition of Protein Deubiquitination by PR-619 Activates the Autophagic Pathway in OLN-t40 Oligodendroglial Cells

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Abstract

Protein aggregate formation may be the result of an impairment of the protein quality control system, e.g., the ubiquitin proteasome system (UPS) and the lysosomal autophagic pathway. For proteasomal degradation, proteins need to be covalently modified by ubiquitin and deubiquitinated before the substrates are proteolytically degraded. Deubiquitination is performed by a large family of proteases, the deubiquitinating enzymes (DUBs). DUBs display a variety of functions and their inhibition may have pathological consequences. Using the broad specificity DUB inhibitor PR-619 we previously have shown that DUB inhibition leads to an overload of ubiquitinated proteins, to protein aggregate formation and subsequent inhibition of the UPS. This study was undertaken to investigate whether PR-619 modulates autophagic functions to possibly compensate the failure of the proteasomal system. Using the oligodendroglial cell line OLN-t40 and a new oligodendroglial cell line stably expressing GFP-LC3, we show that DUB inhibition leads to the activation of autophagy and to the recruitment of LC3 and of the ubiquitin binding protein p62 to the forming aggresomes without impairing the autophagic flux. Furthermore, PR-619 induced the transport of lysosomes to the forming aggregates in a process requiring an intact microtubule network. Further stimulation of autophagy by rapamycin did not prevent PR-619 aggregate formation but rather exerted cytotoxic effects. Hence, inhibition of DUBs by PR-619 activated the autophagic pathway supporting the hypothesis that the UPS and the autophagy–lysosomal pathway are closely linked together.

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Acknowledgments

This study was supported by the Deutsche Forschungsgemeinschaft. The expert technical help of Angelika Spanjer is gratefully acknowledged. We thank Dr. Olaf Goldbaum for helpful discussions.

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Authors and Affiliations

  1. Department of Biology, Molecular Neurobiology, University of Oldenburg, POB 2503, 26111, Oldenburg, Germany

    Veronika Seiberlich, Janika Borchert & Christiane Richter-Landsberg

  2. LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA, 19355, USA

    Victoria Zhukareva

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  1. Veronika Seiberlich
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  2. Janika Borchert
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  3. Victoria Zhukareva
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  4. Christiane Richter-Landsberg
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Correspondence to Victoria Zhukareva or Christiane Richter-Landsberg.

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Seiberlich, V., Borchert, J., Zhukareva, V. et al. Inhibition of Protein Deubiquitination by PR-619 Activates the Autophagic Pathway in OLN-t40 Oligodendroglial Cells. Cell Biochem Biophys 67, 149–160 (2013). https://doi.org/10.1007/s12013-013-9622-8

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