Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis

Petrie, Emma J.; Sandow, Jarrod J.; Jacobsen, Annette V.; Smith, Brian J.; Griffin, Michael D. W.; Lucet, Isabelle S; Dai, Wayne Wei-Ming; Young, Samuel N.; Tanzer, Maria C.; Wardak, Ahmad; Liang, Lung-Yu; Cowan, A.D.; Hildebrand, Joanne M; Kersten, Wilhelmus J A; Lessène, Guillaume; Silke, John; Czabotar, P.E.; Webb, Andrew I.; Murphy, James M.

doi:10.1038/s41467-018-04714-7

Cited by 167 publications

(298 citation statements)

References 47 publications

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“…The Glu351Lys mutation of human MLKL, which was discovered to lead to enhancement of ATP binding, disrupts the formation of MLKL tetramers and thus subsiding octamers. On the contrary, phosphomimic mutations at Thr357 and Ser358 in the human MLKL stabilize the tetramers, indicating that phosphorylation of MLKL protects from ATP-induced dissociation of oligomers [68]. However, human MLKL with mutations of Lys230Met/Gln356Ala in its ATP-binding pocket can still activate itself and translocate to the plasma membrane to induce necroptosis [69].…”

Section: Mlkl Is the Effector Protein In Necroptosismentioning

confidence: 99%

Molecular Insights into the Mechanism of Necroptosis: The Necrosome as a Potential Therapeutic Target

Chen

Garssen

et al. 2019

Cells

134

115

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Necroptosis, or regulated necrosis, is an important type of programmed cell death in addition to apoptosis. Necroptosis induction leads to cell membrane disruption, inflammation and vascularization. It plays important roles in various pathological processes, including neurodegeneration, inflammatory diseases, multiple cancers, and kidney injury. The molecular regulation of necroptotic pathway has been intensively studied in recent years. Necroptosis can be triggered by multiple stimuli and this pathway is regulated through activation of receptor-interacting protein kinase 1 (RIPK1), RIPK3 and pseudokinase mixed lineage kinase domain-like (MLKL). A better understanding of the mechanism of regulation of necroptosis will further aid to the development of novel drugs for necroptosis-associated human diseases. In this review, we focus on new insights in the regulatory machinery of necroptosis. We further discuss the role of necroptosis in different pathologies, its potential as a therapeutic target and the current status of clinical development of drugs interfering in the necroptotic pathway.

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Section: Mlkl Is the Effector Protein In Necroptosismentioning

confidence: 99%

Molecular Insights into the Mechanism of Necroptosis: The Necrosome as a Potential Therapeutic Target

Chen

Garssen

et al. 2019

Cells

134

115

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“…It is well established that RIPK3-mediated phosphorylation of MLKL (at mouse S345 and human T357/S358) is a hallmark of necroptotic signaling Rodriguez et al, 2016;Sun et al, 2012) and an essential signaling event in the pathway owing to its role in promoting assembly of killer MLKL oligomers. The precise role of phosphorylation differs between mouse and human MLKL activation Petrie et al, 2018Petrie et al, , 2019Tanzer et al, 2015); phosphorylation of MLKL in mouse cells occurs via a transient ''kiss and run'' mechanism (Hildebrand et al, 2014;Murphy et al, 2013), while in human cells, phosphorylation of MLKL occurs after stable recruitment to the necrosome via the RIPK3 kinase domain Sun et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Viral MLKL Homologs Subvert Necroptotic Cell Death by Sequestering Cellular RIPK3

et al. 2019

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Necroptotic cell death has been implicated in many human pathologies and is thought to have evolved as an innate immunity mechanism. The pathway relies on two key effectors: the kinase receptor-interacting protein kinase 3 (RIPK3) and the terminal effector, the pseudokinase mixed-lineage kinasedomain-like (MLKL). We identify proteins with high sequence similarity to the pseudokinase domain of MLKL in poxvirus genomes. Expression of these proteins from the BeAn 58058 and Cotia poxviruses, but not swinepox, in human and mouse cells blocks cellular MLKL activation and necroptotic cell death. We show that viral MLKL-like proteins function as dominant-negative mimics of host MLKL, which inhibit necroptosis by sequestering RIPK3 via its kinase domain to thwart MLKL engagement and phosphorylation. These data support an ancestral role for necroptosis in defense against pathogens. Furthermore, mimicry of a cellular pseudokinase by a pathogen adds to the growing repertoire of functions performed by pseudokinases in signal transduction.

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“…It is interesting to note that in human MLKL, no helical insert has been reported. These data suggest that α C and the activation loop helix may serve pseudokinase specific functions that may even differ between species as observed for MLKL (Petrie et al, 2018). Our evolutionary analysis suggests that the loss of lysine-glutamate salt bridge in metazoan ULK4 required the stabilization of α C by the activation segment helix which in metazoan ULK4 has co-occurred with an extended activation loop, presumably compensating for the loss of the canonical K-E salt bridge.…”

Section: Degradation Of the Active Site Has Co-evolved With An Extendmentioning

confidence: 61%

Nucleotide binding, evolutionary insights and interaction partners of the pseudokinase Unc-51-like kinase 4

Preuß

Chatterjee

Mathea

et al. 2020

Preprint

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Unc-51-like kinase 4 (ULK4) is a pseudokinase that has been linked to the development of several diseases. Even though sequence motifs required for ATP binding in kinases are lacking, ULK4 still tightly binds ATP and the presence of the cofactor is required for structural stability of ULK4. Here we present a high-resolution structure of a ULK4-ATPγS complex revealing a highly unusual ATP binding mode in which the lack of the canonical VAIK motif lysine is compensated by K39, located N-terminal to αC. Evolutionary analysis suggests that degradation of active site motifs in metazoan ULK4 has co-occurred with an ULK4 specific activation loop, which stabilizes the C-helix. In addition, cellular interaction studies using BioID and biochemical validation data revealed high confidence interactors of the pseudokinase and armadillo repeat domains. Many of the identified ULK4 interaction partners were centrosomal and tubulin associated proteins and several active kinases suggesting new roles for ULK4. Highlights:Structure of the ULK4 ATP complex reveals a unique ATP binding mode. Disease associated mutations modulate ATP binding and ULK4 stabilityDegradation of active site motifs co-occurred in evolution with an ULK4 specific activation loop BioID suggests a role of ULK4 regulating centrosomal and cytoskeletal functions

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Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis

Cited by 167 publications

References 47 publications

Molecular Insights into the Mechanism of Necroptosis: The Necrosome as a Potential Therapeutic Target

Molecular Insights into the Mechanism of Necroptosis: The Necrosome as a Potential Therapeutic Target

Viral MLKL Homologs Subvert Necroptotic Cell Death by Sequestering Cellular RIPK3

Nucleotide binding, evolutionary insights and interaction partners of the pseudokinase Unc-51-like kinase 4

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