Nuclear inclusions of pathogenic ataxin-1 induce oxidative stress and perturb the protein synthesis machinery
Stamatia Laidou,
Gregorio Alanis-Lobato,
Jan Pribyl,
Tamás Raskó,
Boris Tichy,
Kamil Mikulasek,
Maria Tsagiopoulou,
Jan Oppelt,
Georgia Kastrinaki,
Maria Lefaki,
Manvendra Singh,
Annika Zink,
Niki Chondrogianni,
Fotis Psomopoulos,
Alessandro Prigione,
Zoltán Ivics,
Sarka Pospisilova,
Petr Skladal,
Zsuzsanna Izsvák,
Miguel A. Andrade-Navarro,
Spyros Petrakis
Affiliations
Stamatia Laidou
Institute of Applied Biosciences/Centre for Research and Technology Hellas, 57001, Thessaloniki, Greece
Gregorio Alanis-Lobato
Faculty of Biology, Johannes Gutenberg University Mainz, 55122, Mainz, Germany; Human Embryo and Stem Cell Laboratory, The Francis Crick Institute, NW1 1AT, London, UK
Jan Pribyl
Central European Institute of Technology, Masaryk University, 62500, Brno, Czech Republic
Tamás Raskó
Max-Delbrueck-Center for Molecular Medicine in the Helmholtz Association, Berlin, 13125, Germany
Boris Tichy
Central European Institute of Technology, Masaryk University, 62500, Brno, Czech Republic
Kamil Mikulasek
Central European Institute of Technology, Masaryk University, 62500, Brno, Czech Republic; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, 62500, Brno, Czech Republic
Maria Tsagiopoulou
Institute of Applied Biosciences/Centre for Research and Technology Hellas, 57001, Thessaloniki, Greece
Jan Oppelt
Central European Institute of Technology, Masaryk University, 62500, Brno, Czech Republic
Georgia Kastrinaki
Aerosol and Particle Technology Laboratory/Chemical Process & Energy Resources Institute/Centre for Research and Technology Hellas, 57001, Thessaloniki, Greece
Maria Lefaki
Institute of Biology, Medicinal Chemistry & Biotechnology/National Hellenic Research Foundation, 11365, Athens, Greece
Manvendra Singh
Max-Delbrueck-Center for Molecular Medicine in the Helmholtz Association, Berlin, 13125, Germany
Annika Zink
Max-Delbrueck-Center for Molecular Medicine in the Helmholtz Association, Berlin, 13125, Germany; Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich Heine University, 40225, Düsseldorf, Germany
Niki Chondrogianni
Institute of Biology, Medicinal Chemistry & Biotechnology/National Hellenic Research Foundation, 11365, Athens, Greece
Fotis Psomopoulos
Institute of Applied Biosciences/Centre for Research and Technology Hellas, 57001, Thessaloniki, Greece; Department of Molecular Medicine and Surgery, Karolinska Institutet, 17177, Stockholm, Sweden
Alessandro Prigione
Max-Delbrueck-Center for Molecular Medicine in the Helmholtz Association, Berlin, 13125, Germany; Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich Heine University, 40225, Düsseldorf, Germany
Zoltán Ivics
Division of Medical Biotechnology, Paul-Ehrlich-Institute, 63225, Langen, Germany
Sarka Pospisilova
Central European Institute of Technology, Masaryk University, 62500, Brno, Czech Republic
Petr Skladal
Central European Institute of Technology, Masaryk University, 62500, Brno, Czech Republic
Zsuzsanna Izsvák
Max-Delbrueck-Center for Molecular Medicine in the Helmholtz Association, Berlin, 13125, Germany; Corresponding author.
Miguel A. Andrade-Navarro
Faculty of Biology, Johannes Gutenberg University Mainz, 55122, Mainz, Germany
Spyros Petrakis
Institute of Applied Biosciences/Centre for Research and Technology Hellas, 57001, Thessaloniki, Greece; Corresponding author.
Spinocerebellar ataxia type-1 (SCA1) is caused by an abnormally expanded polyglutamine (polyQ) tract in ataxin-1. These expansions are responsible for protein misfolding and self-assembly into intranuclear inclusion bodies (IIBs) that are somehow linked to neuronal death. However, owing to lack of a suitable cellular model, the downstream consequences of IIB formation are yet to be resolved. Here, we describe a nuclear protein aggregation model of pathogenic human ataxin-1 and characterize IIB effects. Using an inducible Sleeping Beauty transposon system, we overexpressed the ATXN1(Q82) gene in human mesenchymal stem cells that are resistant to the early cytotoxic effects caused by the expression of the mutant protein. We characterized the structure and the protein composition of insoluble polyQ IIBs which gradually occupy the nuclei and are responsible for the generation of reactive oxygen species. In response to their formation, our transcriptome analysis reveals a cerebellum-specific perturbed protein interaction network, primarily affecting protein synthesis. We propose that insoluble polyQ IIBs cause oxidative and nucleolar stress and affect the assembly of the ribosome by capturing or down-regulating essential components. The inducible cell system can be utilized to decipher the cellular consequences of polyQ protein aggregation. Our strategy provides a broadly applicable methodology for studying polyQ diseases.