The Pseudokinase MLKL Mediates Necroptosis via a Molecular Switch Mechanism
Open accessAbstract
Mixed lineage kinase domain-like (MLKL) is a component of the "necrosome," the multiprotein complex that triggers tumor necrosis factor (TNF)-induced cell death by necroptosis. To define the specific role and molecular mechanism of MLKL action, we generated MLKL-deficient mice and solved the crystal structure of MLKL. Although MLKL-deficient mice were viable and displayed no hematopoietic anomalies or other obvious pathology, cells derived from these animals were resistant to TNF-induced necroptosis unless MLKL expression was restored. Structurally, MLKL comprises a four-helical bundle tethered to the pseudokinase domain, which contains an unusual pseudoactive site. Although the pseudokinase domain binds ATP, it is catalytically inactive and its essential nonenzymatic role in necroptotic signaling is induced by receptor-interacting serine-threonine kinase 3 (RIPK3)-mediated phosphorylation. Structure-guided mutation of the MLKL pseudoactive site resulted in constitutive, RIPK3-independent necroptosis, demonstrating that modification of MLKL is essential for propagation of the necroptosis pathway downstream of RIPK3.
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Necroptosis critically depends on sequential activation of RIPK3 and MLKL.
“At the molecular level, necroptosis critically depends on the sequential activation of RIPK3 and mixed lineage kinase domain like pseudokinase (MLKL) [434, 435].”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [435]
Essential contribution of MLKL to necroptosis has been confirmed by genetic studies and pharmacological interventions.
“Although the essential contribution of MLKL to necroptosis has been confirmed by genetic studies [435] as well as by pharmacological (i.e., inhibition of MLKL with necrosulfonamide, NSA) interventions [442]”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [435]
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- Authors
- Murphy, J. M., Czabotar, P. E., Hildebrand, J. M., Lucet, I. S., Zhang, J.-G., Alvarez-Diaz, S., Lewis, R., Lalaoui, N., Metcalf, D., Webb, A. I., Young, S. N., Varghese, L. N., Tannahill, G. M., Hatchell, E. C., Majewski, I. J., Okamoto, T., Dobson, R. C. J., Hilton, D. J., Babon, J. J., Nicola, N. A., Strasser, A., Silke, J., & Alexander, W. S.
- Affiliations
- The University of Melbourne (AU)
- Monash University (AU)
- Corresponding author
- Murphy, James M.
- ORCID
- Murphy, James M.
- Czabotar, Peter E.
- Hildebrand, Joanne M.
- Lucet, Isabelle S.
- Zhang, Jian-Guo
- Lalaoui, Najoua
- Webb, Andrew I.
- Young, Samuel N.
- Varghese, Leila N.
- Majewski, Ian J.
- Okamoto, Toru
- Dobson, Renwick C.J.
- Hilton, Douglas J.
- Babon, Jeffrey J.
- Nicola, Nicos A.
- Strasser, Andreas
- Silke, John
- Alexander, Warren S.
- Venue
- Immunity
- Publisher
- Elsevier BV
- Published
- 2013-09
- Volume
- 39
- Issue
- 3
- Pages
- 443-453
- ISSN
- 1074-7613 (ISSN-L)
- OpenAlex
- W1993613360
- Cited by
- 1,220
- References
- 3
- Citations by year
- 2026: 67 · 2025: 112 · 2024: 115 · 2023: 94 · 2022: 113 · 2021: 116 · 2020: 109 · 2019: 89 · 2018: 82 · 2017: 77 · 2016: 91 · 2015: 68 · 2014: 75 · 2013: 12
- FWCI
- 36.76
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Immunity (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Australian Synchrotron
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Keywords
- MLKL, necroptosis, necrosome, RIPK3, TNF-induced cell death, pseudokinase, protein crystal structure, phosphorylation
- MeSH terms
- Animals, Cell Line, Mice, Inbred C57BL, Necrosis, Phosphoric Monoester Hydrolases, Phosphoprotein Phosphatases, Phosphorylation, Protein Kinases, Signal Transduction, Apoptosis, Mice, Knockout, Crystallography, X-Ray, Catalytic Domain, Tumor Necrosis Factors, Mice, Receptor-Interacting Protein Serine-Threonine Kinases
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