2. Academic Validation
  3. The deubiquitylase USP9X controls ribosomal stalling

The deubiquitylase USP9X controls ribosomal stalling

  • J Cell Biol. 2021 Mar 1;220(3):e202004211. doi: 10.1083/jcb.202004211.
Anne Clancy 1,  Claire Heride  # 1 2,  Adán Pinto-Fernández  # 3,  Hannah Elcocks 1,  Andreas Kallinos 1,  Katherine J Kayser-Bricker 4,  Weiping Wang 4,  Victoria Smith 1,  Simon Davis 3,  Shawn Fessler 4,  Crystal McKinnon 4,  Marie Katz 4,  Tim Hammonds 2,  Neil P Jones 2,  Jonathan O'Connell 4,  Bruce Follows 4,  Steven Mischke 4,  Justin A Caravella 4,  Stephanos Ioannidis 4,  Christopher Dinsmore 4,  Sunkyu Kim 4,  Axel Behrens 5,  David Komander 6 7,  Benedikt M Kessler 3,  Sylvie Urbé 1,  Michael J Clague 1
Affiliations

Affiliations

  • 1 Department of Molecular Physiology and Cell Signaling, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, UK.
  • 2 Cancer Research UK Therapeutic Discovery Laboratories, London Bioscience Innovation Centre, London, UK.
  • 3 Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
  • 4 FORMA Therapeutics, Watertown, MA.
  • 5 Adult Stem Cell Laboratory, Francis Crick Institute, London, UK.
  • 6 Ubiquitin Signalling Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
  • 7 Department of Medical Biology, University of Melbourne, Melbourne, Victoria, Australia.
  • # Contributed equally.
PMID: 33507233 DOI: 10.1083/jcb.202004211
Abstract

When a ribosome stalls during translation, it runs the risk of collision with a trailing ribosome. Such an encounter leads to the formation of a stable di-ribosome complex, which needs to be resolved by a dedicated machinery. The initial stalling and the subsequent resolution of di-ribosomal complexes requires activity of Makorin and ZNF598 ubiquitin E3 Ligases, respectively, through ubiquitylation of the eS10 and uS10 subunits of the ribosome. We have developed a specific small-molecule inhibitor of the deubiquitylase USP9X. Proteomics analysis, following inhibitor treatment of HCT116 cells, confirms previous reports linking USP9X with centrosome-associated protein stability but also reveals a loss of Makorin 2 and ZNF598. We show that USP9X interacts with both these ubiquitin E3 Ligases, regulating their abundance through the control of protein stability. In the absence of USP9X or following chemical inhibition of its catalytic activity, levels of Makorins and ZNF598 are diminished, and the ribosomal quality control pathway is impaired.

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