MLKL activity requires a splicing-regulated, druggable intramolecular interaction
Necroptosis is an inflammatory form of regulated cell death implicated in a range of human pathologies, whose execution depends on the poorly understood pseudokinase mixed lineage kinase domain-like (MLKL). Here, we report that splicing-dependent insertion of a short amino acid sequence in the C-terminal α-helix (Hc) of MLKL abolishes cell killing activity and creates an anti-necroptotic isoform that counteracts cell death induced by the necroptosis-proficient protein in mice and humans. We show that interaction of Hc with a previously unrecognized hydrophobic groove is essential for necroptosis, which we exploited in a strategy to identify small molecules that inhibit MLKL and substantially ameliorate disease in murine models of necroptosis-driven dermatitis and abdominal aortic aneurysm. Thus, alternative splicing of microexons controls the ability of MLKL to undergo an intramolecular rearrangement essential for necroptosis with potential to guide the development of allosteric MLKL inhibitors for the treatment of human disease.
Authors
- D. Peter Tieleman (ORCID: https://orcid.org/0000-0001-5507-0688)
- Harshal Nemade (ORCID: https://orcid.org/0000-0002-2704-181X)
- J. A. Weir
- Uris Ros (ORCID: https://orcid.org/0000-0002-8889-0145)
- Miloš Gojković (ORCID: https://orcid.org/0000-0001-6451-9192)
- Rafael A. Espiritu (ORCID: https://orcid.org/0000-0001-9993-4310)
- Nieves Peltzer (ORCID: https://orcid.org/0000-0002-4134-5852)
- Julia Saggau (ORCID: https://orcid.org/0000-0002-9908-5467)
- Julian D. Langer (ORCID: https://orcid.org/0000-0002-5190-577X)
- W. Wei‐Lynn Wong (ORCID: https://orcid.org/0000-0003-3155-1211)
- Ana J. García‐Sáez (ORCID: https://orcid.org/0000-0002-3894-5945)
- Martin Mollenhauer (ORCID: https://orcid.org/0000-0002-3220-9269)
- Hannah Sophia Lottermoser (ORCID: https://orcid.org/0000-0001-8520-2645)
- Veronika Altmannová
- Lazaros Vasilikos (ORCID: https://orcid.org/0000-0003-4071-7025)
- Gianmaria Liccardi (ORCID: https://orcid.org/0000-0002-2662-1281)
- Stefan Müller (ORCID: https://orcid.org/0000-0003-3158-3002)
- Raed Shalaby
- Manolis Pasparakis (ORCID: https://orcid.org/0000-0002-9870-0966)
- Laurens Wachsmuth (ORCID: https://orcid.org/0000-0003-1701-9384)
- Henning Walczak (ORCID: https://orcid.org/0000-0002-6312-4591)
- Matti Adam (ORCID: https://orcid.org/0000-0002-6990-8135)
- Veronica Martinez-Osorio
- Konstantinos Kelepouras (ORCID: https://orcid.org/0000-0003-4257-9404)
- Yuguang Chen (ORCID: https://orcid.org/0009-0007-5915-2358)
- Pedro A. Valiente
- M. K. A. IBRAHIM
- Yasmin Abdelwahab
- J. R. ZOELLER
- Paula Bedoya
- Silvia Zanna
Institutions
- University of Zurich (CH)
- University of Havana (CU)
- University of Tübingen (DE)
Publication Details
- Journal
- Molecular Cell
- Published
- 2025-04-01
- DOI
- https://doi.org/10.1016/j.molcel.2025.03.015
- Citations
- 7
- Primary Topic
- Melanoma and MAPK Pathways
- Type
- article
- Field-Weighted Citation Impact
- 5.64
Funders
- Alexander von Humboldt-Stiftung
- Wellcome Trust
- Deutsche Forschungsgemeinschaft
- Canada Research Chairs
- Bundesministerium für Bildung und Forschung
- Max-Planck-Gesellschaft
- Natural Sciences and Engineering Research Council of Canada
- Medical Research Council
- European Research Council