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Protein products of nonstop mRNA disrupt nucleolar homeostasis

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Cell Stress and Chaperones Aims and scope

Abstract

Stalled mRNA translation results in the production of incompletely synthesized proteins that are targeted for degradation by ribosome-associated quality control (RQC). Here we investigated the fate of defective proteins translated from stall-inducing, nonstop mRNA that escape ubiquitylation by the RQC protein LTN1. We found that nonstop protein products accumulated in nucleoli and this localization was driven by polylysine tracts produced by translation of the poly(A) tails of nonstop mRNA. Nucleolar sequestration increased the solubility of invading proteins but disrupted nucleoli, altering their dynamics, morphology, and resistance to stress in cell culture and intact flies. Our work elucidates how stalled translation may affect distal cellular processes and may inform studies on the pathology of diseases caused by failures in RQC and characterized by nucleolar stress.

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Acknowledgments

We thank the members of the Brandman laboratory for helpful discussions and commentary on the manuscript. Additionally, we acknowledge the members of the Julie Theriot Lab (Stanford/University of Washington) and Aaron Straight (Stanford) labs for use of microscopes, reagents, and expertise. We also thank people from the CIGS microscopy facility (University of Modena and Reggio Emilia) for technical support. This work was performed while ASH was a postdoctoral fellow in the laboratory of Dr. Rohit V. Pappu (RVP) at Washington University in St. Louis and supported by the Human Frontiers Science Program (grant RGP0034/2017 to RVP and S.A.). S.A. was supported by the European Researchn Council (PhaseAge, No. 725836). S.C. acknowledges funding from AriSLA Foundation (MLOpathy); Cariplo Foundation (Rif. 2014-0703); MAECI (Dissolve_ALS); MIUR (Departments of excellence 2018-2022; E91I18001480001). S.C. and S.A. are grateful to EU Joint Programme - Neurodegenerative Disease Research (JPND) project. The project is supported through funding organizations under the aegis of JPND (http://www.neurodegenerationresearch.eu/). This project has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement no. 643417. A preprint of this work was uploaded to BioRxiv (Davis et al., 2019)

Funding

This research was supported by a Stanford Dean’s Postdoctoral Fellowship to ZHD as well as a grant (R01GM115968) from the National Institutes of Health to OB. Work in BL’s lab was supported by grants (R01NS084412, R01NS083417 and R01AR0748750) from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Biochemistry, Stanford University, Stanford, CA, 94305, USA

    Zoe H. Davis & Onn Brandman

  2. Centre for Neuroscience and Nanotechnology, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio, Emilia, Modena, Italy

    Laura Mediani, Francesco Antoniani, Jonathan Vinet & Serena Carra

  3. Department of Pathology, Stanford University, Stanford, CA, 94305, USA

    Shuangxi Li & Bingwei Lu

  4. Biotechnology Center (BIOTEC), Center for Molecular and Cellular Bioengineering (CMCB), Technische Universitat Dresden, Tatzberg 47/49, 01307, Dresden, Germany

    Simon Alberti

  5. Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, 63110, USA

    Alex S. Holehouse

  6. Center for Science and Engineering of Living Systems (CSELS), Washington University in St. Louis, St. Louis, MO, 63130, USA

    Alex S. Holehouse

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  1. Zoe H. Davis
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  2. Laura Mediani
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Correspondence to Serena Carra or Onn Brandman.

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Davis, Z.H., Mediani, L., Antoniani, F. et al. Protein products of nonstop mRNA disrupt nucleolar homeostasis. Cell Stress and Chaperones 26, 549–561 (2021). https://doi.org/10.1007/s12192-021-01200-w

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