Breaking β-sheets in FUS prion-like domain preserves phase separation and function but prevents aggregation and toxicity
The RNA-binding protein Fused in Sarcoma (FUS) undergoes phase separation associated with RNA processing. However, the prion-like low complexity (LC) domain of FUS forms solid-like aggregates in neurodegenerative diseases. Whether the formation of β-sheet structure associated with pathology is also physiologically/functionally relevant is debated. Similarly, if mislocalization alone or concomitant aggregation is responsible for FUS gain-of-function toxicity remains to be probed. Here, we introduce β-sheet breaking proline residues into FUS LC with the goal of preventing cross-β-driven aggregation without disrupting essential functions and phase separation. β-sheet-deficient FUS variants maintain native-like global motions, disorder, and phase separation, but no longer show a liquid-to-solid transition (LST). Biochemical partitioning, cellular localization, and auto- and cross-regulatory functions of FUS all remain essentially unchanged. Conversely, FUS-induced neurodegeneration in several <i>Drosophila</i> models is drastically reduced. These findings suggest a strategy for mitigating disease-related toxicity through backbone structure modulation to prevent prion-like domain protein aggregation.
Authors
- Daniel Jutzi (ORCID: https://orcid.org/0000-0001-8381-7830)
- S. Wang
- Noah Wake (ORCID: https://orcid.org/0000-0003-4119-2574)
- Benjamin S. Schuster (ORCID: https://orcid.org/0000-0002-6468-8081)
- Mayur Barai
- Marc‐David Ruepp (ORCID: https://orcid.org/0000-0003-3264-9800)
- Eric N. Anderson (ORCID: https://orcid.org/0000-0003-2533-8764)
- Udai Bhan Pandey (ORCID: https://orcid.org/0000-0002-6267-0179)
- Nicolas Lux Fawzi (ORCID: https://orcid.org/0000-0001-5483-0577)
- Ryan Z. Puterbaugh (ORCID: https://orcid.org/0000-0002-6412-8703)
- Shuo-Lin Weng (ORCID: https://orcid.org/0009-0009-8082-8206)
- S R Cummings (ORCID: https://orcid.org/0009-0003-7605-4099)
- Tongyin Zheng
- Daryl A. Bosco
- Juan Alcalde
- Jeetain Mittal
- Anjali Bajaj
- Sukleen Kour
Institutions
- Rutgers, The State University of New Jersey (US)
- UMass Memorial Health Care (US)
- University of Massachusetts Chan Medical School (US)
- King's College London (GB)
- Brown University (US)
- Environmental and Occupational Health Sciences Institute (US)
- UK Dementia Research Institute (GB)
- Children's Hospital of Pittsburgh (US)
- University of Pittsburgh Medical Center (US)
- UMass Memorial Medical Center (US)
- Texas A&M University (US)
- University of Edinburgh (GB)
Publication Details
- Journal
- bioRxiv (Cold Spring Harbor Laboratory)
- Published
- 2026-02-18
- DOI
- https://doi.org/10.64898/2026.02.17.706410
- Citations
- 1
- Primary Topic
- RNA Research and Splicing
- Type
- article
- Field-Weighted Citation Impact
- 22.24
Funders
- Brown University
- Motor Neurone Disease Association
- King's College London
- NOMIS Stiftung
- UK Dementia Research Institute
- High Performance Research Computing, Texas A and M University
- Medical Research Council
- National Institute of General Medical Sciences