title: Inhibition of protein deubiquitination by PR-619 activates the autophagic pathway in OLN-t40 oligodendroglial cells.
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.
authors.affiliation: Department of Biology, Molecular Neurobiology, University of Oldenburg, POB 2503, 26111, Oldenburg, Germany.
authors.forename: Veronika
authors.initials: V
authors.lastname: Seiberlich
authors.forename: Janika
authors.initials: J
authors.lastname: Borchert
authors.forename: Victoria
authors.initials: V
authors.lastname: Zhukareva
authors.forename: Christiane
authors.initials: C
authors.lastname: Richter-Landsberg
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000631:Aminopyridines / Q000737:chemistry / Q000633:toxicity*; D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry / Q000633:toxicity*; D051381:Rats; D000071456:Sequestosome-1 Protein; D020123:Sirolimus / Q000633:toxicity; D013861:Thiocyanates / Q000737:chemistry / Q000633:toxicity*; D064570:Ubiquitin-Specific Proteases / Q000737:chemistry / Q000378:metabolism*; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000631:Aminopyridines; D008869:Microtubule-Associated Proteins; D011480:Protease Inhibitors; D000071456:Sequestosome-1 Protein; D020123:Sirolimus; D013861:Thiocyanates; D064570:Ubiquitin-Specific Proteases; D016875:tau Proteins; C570894:2,6-diaminopyridine-3,5-bis(thiocyanate); C526109:Anxa3 protein, rat; C099718:SQSTM1 protein, human