The Pseudokinase MLKL Mediates Necroptosis via a Molecular Switch Mechanism

Murphy, James M.; Czabotar, P.E.; Hildebrand, Joanne M; Lucet, Isabelle S; Zhang, Jianguo; Álvarez-Díaz, Silvia; Lewis, Rowena S.; Lalaoui, Najoua; Metcalf, Donald; Webb, Andrew I.; Young, Samuel N.; Varghese, Leila N.; Tannahill, Gillian M.; Hatchell, Esme C.; Majewski, Ian J.; Okamoto, Tomoyoshi; Dobson, Renwick C. J.; Hilton, Douglas J.; Babon, Jeffrey J.; Nicola, Nicos A.; Strasser, Andreas; Silke, John; Alexander, Warren S.

doi:10.1016/j.immuni.2013.06.018

Cited by 1,041 publications

(1,311 citation statements)

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“…Necroptosis involves the formation of the necrosome, a complex consisting of RIP (receptor serine–threonine–protein kinase)1, RIP3 and MLKL (mixed lineage kinase domain‐like) and similarly to pyroptosis it is characterized by cell lysis (Pasparakis & Vandenabeele, 2015). Recent work has demonstrated that recruitment of MLKL to RIP3 and MLKL phosphorylation results in the conformational change that leads to the exposure of its four‐helix bundle domain (FHBD) and oligomerization of MLKL (Murphy et al , 2013). Binding of the FHBD to negatively charged phosphatidylinositol phosphates has been proposed to recruit MLKL to the plasma membrane allowing it to directly permeabilize the membrane by forming a pore (Dondelinger et al , 2014; Su et al , 2014; Wang et al , 2014).…”

Section: Discussionmentioning

confidence: 99%

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

et al. 2016

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Pyroptosis is a lytic type of cell death that is initiated by inflammatory caspases. These caspases are activated within multi‐protein inflammasome complexes that assemble in response to pathogens and endogenous danger signals. Pyroptotic cell death has been proposed to proceed via the formation of a plasma membrane pore, but the underlying molecular mechanism has remained unclear. Recently, gasdermin D (GSDMD), a member of the ill‐characterized gasdermin protein family, was identified as a caspase substrate and an essential mediator of pyroptosis. GSDMD is thus a candidate for pyroptotic pore formation. Here, we characterize GSDMD function in live cells and in vitro. We show that the N‐terminal fragment of caspase‐1‐cleaved GSDMD rapidly targets the membrane fraction of macrophages and that it induces the formation of a plasma membrane pore. In vitro, the N‐terminal fragment of caspase‐1‐cleaved recombinant GSDMD tightly binds liposomes and forms large permeability pores. Visualization of liposome‐inserted GSDMD at nanometer resolution by cryo‐electron and atomic force microscopy shows circular pores with variable ring diameters around 20 nm. Overall, these data demonstrate that GSDMD is the direct and final executor of pyroptotic cell death.

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Section: Discussionmentioning

confidence: 99%

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

et al. 2016

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“…However, a more recent study has questioned the role of PGAM5 in mediating TNF-induced necrosis, at least in murine fibroblasts. 87 Knockdown of PGAM5 expression in these cells failed to affect susceptibility to TNF-driven necroptosis, suggesting alternative or additional mediatory pathways. Such discrepancies may reflect varying effector mechanisms in different cells and species.…”

Section: Rip1 and Rip3 As Drivers Of Necroptosismentioning

confidence: 91%

“…84 The recruitment of MLKL to the necrosome leads to RIP3-mediated phosphorylation of MLKL with the RIP3 inhibitor, necrosulfonamide, blocking necrosis downstream of RIP3 activation 85 and MLKL-deficient mice being resistant to necroptosis. 86,87 Various downstream effector mechanisms have been proposed to mediate necroptosis. Phosphorylation of MLKL promotes its oligomerization and translocation to the plasma membrane where it interacts with phospholipids and compromises membrane integrity ultimately resulting in cell rupture.…”

Section: Rip1 and Rip3 As Drivers Of Necroptosismentioning

confidence: 99%

RIP kinases: key decision makers in cell death and innate immunity

et al. 2014

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“…[31][32][33][34] Necrosome also suppresses apoptosis but the underlying mechanism has not been described yet. Mixed-lineage kinase domain-like (MLKL) is downstream of RIP3, 35,36 and phosphorylation of MLKL is required for necroptosis. [37][38][39][40][41][42] Apoptosis inducing complex (complex II) and necrosome are both supramolecular complexes.…”

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confidence: 99%

Distinct roles of RIP1–RIP3 hetero- and RIP3–RIP3 homo-interaction in mediating necroptosis

et al. 2014

Cell Death Differ

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Necroptosis is mediated by a signaling complex called necrosome, containing receptor-interacting protein (RIP)1, RIP3, and mixed-lineage kinase domain-like (MLKL). It is known that RIP1 and RIP3 form heterodimeric filamentous scaffold in necrosomes through their RIP homotypic interaction motif (RHIM) domain-mediated oligomerization, but the signaling events based on this scaffold has not been fully addressed. By using inducible dimer systems we found that RIP1-RIP1 interaction is dispensable for necroptosis; RIP1-RIP3 interaction is required for necroptosis signaling, but there is no necroptosis if no additional RIP3 protein is recruited to the RIP1-RIP3 heterodimer, and the interaction with RIP1 promotes the RIP3 to recruit other RIP3; RIP3-RIP3 interaction is required for necroptosis and RIP3-RIP3 dimerization is sufficient to induce necroptosis; and RIP3 dimer-induced necroptosis requires MLKL. We further show that RIP3 oligomer is not more potent than RIP3 dimer in triggering necroptosis, suggesting that RIP3 homo-interaction in the complex, rather than whether RIP3 has formed homo polymer, is important for necroptosis. RIP3 dimerization leads to RIP3 intramolecule autophosphorylation, which is required for the recruitment of MLKL. Interestingly, phosphorylation of one of RIP3 in the dimer is sufficient to induce necroptosis. As RIP1-RIP3 heterodimer itself cannot induce necroptosis, the RIP1-RIP3 heterodimeric amyloid fibril is unlikely to directly propagate necroptosis. We propose that the signaling events after the RIP1-RIP3 amyloid complex assembly are the recruitment of free RIP3 by the RIP3 in the amyloid scaffold followed by autophosphorylation of RIP3 and subsequent recruitment of MLKL by RIP3 to execute necroptosis. Cell Death and Differentiation (2014) 21, 1709-1720; doi:10.1038/cdd.2014.77; published online 6 June 2014Necroptosis is a type of programmed necrosis characterized by necrotic morphological changes, including cellular organelle swelling, cell membrane rupture, 1-3 and dependence of receptor-interacting protein (RIP)1 4 and RIP3. [5][6][7] Physiological function of necroptosis has been illustrated in host defense, [8][9][10][11] inflammation, 12-16 tissue injury, 10,17,18 and development. [19][20][21] Necroptosis can be induced by a number of different extracellular stimuli such as tumor necrosis factor (TNF). TNF stimulation leads to formation of TNF receptor 1 (TNFR1) signaling complex (named complex I), and complex II containing RIP1, TRADD, FAS-associated protein with a death domain (FADD), and caspase-8, of which the activation initiates apoptosis. If cells have high level of RIP3, RIP1 recruits RIP3 to form necrosome containing FADD, [22][23][24] caspase-8, RIP1, and RIP3, and the cells undergo necroptosis. 25,26 Caspase-8 and FADD negatively regulates necroptosis, 27-30 because RIP1, RIP3, and CYLD are potential substrates of caspase-8. [31][32][33][34] Necrosome also suppresses apoptosis but the underlying mechanism has not been described yet. Mixed-lineage kinase domain-like (ML...

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The Pseudokinase MLKL Mediates Necroptosis via a Molecular Switch Mechanism

Cited by 1,041 publications

References 43 publications

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

RIP kinases: key decision makers in cell death and innate immunity

Distinct roles of RIP1–RIP3 hetero- and RIP3–RIP3 homo-interaction in mediating necroptosis

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