Sequential Engagement of Distinct MLKL Phosphatidylinositol-Binding Sites Executes Necroptosis

Quarato, Giovanni; Guy, Cliff; Grace, Christy R.; Llambi, Fabien; Nourse, Amanda; Rodríguez, Diego A.; Wakefield, Randall; Frase, Sharon; Moldoveanu, Tudor; Green, Douglas R.

doi:10.1016/j.molcel.2016.01.011

Cited by 199 publications

(305 citation statements)

References 28 publications

Supporting

Mentioning

283

Contrasting

Unclassified

Order By: Relevance

“…The information known about this process is that autophosphorylated RIP3 in the necrosome recruits and phosphorylates MLKL, which then oligomerizes and translocates to the plasma membrane (23,26,34,39). In this study, we show that MLKL actually forms an octamer ( Fig.…”

Section: Discussionmentioning

confidence: 50%

“…MLKL is a well-known effector of necroptosis and is activated downstream of RIP3 (24)(25)(26)(27)(28)(29)(31)(32)(33)(34). Although its monomer structure has been solved and a number of amino acids or clusters of amino acids were revealed to be important for MLKL oligomerization and targeting to the plasma membrane (27,32,(34)(35)(36), the details of its oligomerization and oligomer structure are still largely unknown.…”

mentioning

confidence: 99%

“…Although its monomer structure has been solved and a number of amino acids or clusters of amino acids were revealed to be important for MLKL oligomerization and targeting to the plasma membrane (27,32,(34)(35)(36), the details of its oligomerization and oligomer structure are still largely unknown. Here we show for the first time that MLKL oligomerizes in the necrosome and is then released as the fully assembled octamer.…”

mentioning

confidence: 99%

See 2 more Smart Citations

The MLKL Channel in Necroptosis Is an Octamer Formed by Tetramers in a Dyadic Process

Huang

Zheng

Wang

et al. 2017

Molecular and Cellular Biology

110

102

View full text Add to dashboard Cite

Oligomerization of the mixed-lineage kinase domain-like protein (MLKL) is essential for its cation channel function in necroptosis. Here we show that the MLKL channel is an octamer comprising two previously identified tetramers most likely in their side-by-side position. Intermolecule disulfide bonds are present in the tetramer but are not required for octamer assembly and necroptosis. MLKL forms oligomers in the necrosome and is then released from the necrosome before or during its membrane translocation. We identified two MLKL mutants that could not oligomerize into octamers, although they formed a tetramer, and also, one MLKL mutant could spontaneously form a disulfide bond-linked octamer. Subsequent analysis revealed that the tetramers fail to translocate to the plasma membrane and that the MLKL octamer formation depends on ␣-helices 4 and 5. While MLKL could be detected from outside the cells, its N-and C-terminal ends could not be detected, indicating that the MLKL octamer spans across the plasma membrane, leaving its N and C termini inside the cell. These data allowed us to propose a 180°symmetry model of the MLKL octamer and conclude that the fully assembled MLKL octamers, but not the previously described tetramers, act as effectors of necroptosis.

show abstract

Section: Discussionmentioning

confidence: 50%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

The MLKL Channel in Necroptosis Is an Octamer Formed by Tetramers in a Dyadic Process

Huang

Zheng

Wang

et al. 2017

Molecular and Cellular Biology

110

102

View full text Add to dashboard Cite

show abstract

“…To exclude the possibility that the polymers purified from the cells might still contain other proteins, we turned to a recombinant system. The NTD of MLKL is sufficient to induce necroptosis when overexpressed (20,23,24). Moreover, recombinant NTD could permeabilize liposomes (17,21,22).…”

Section: Mlkl Forms Polymers Upon Necroptosis Induction In Both Humanmentioning

confidence: 99%

“…However, how MLKL oligomers kill cells is still under debate. Recombinant MLKL was shown to bind phospholipids and cause liposome leakage in vitro, leading to the model that oligomerized MLKL could form pores to permeabilize cell membrane (17,(21)(22)(23). Nonetheless, membrane translocation alone is not sufficient for cell death induction, because some MLKL mutants can form oligomers and translocate to the cell membrane but cannot kill cells (24).…”

mentioning

confidence: 99%

MLKL forms disulfide bond-dependent amyloid-like polymers to induce necroptosis

Liu

Johnston

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

136

123

View full text Add to dashboard Cite

Mixed-lineage kinase domain-like protein (MLKL) is essential for TNF-α-induced necroptosis. How MLKL promotes cell death is still under debate. Here we report that MLKL forms SDS-resistant, disulfide bond-dependent polymers during necroptosis in both human and mouse cells. MLKL polymers are independent of receptor-interacting protein kinase 1 and 3 (RIPK1/RIPK3) fibers. Large MLKL polymers are more than 2 million Da and are resistant to proteinase K digestion. MLKL polymers are fibers 5 nm in diameter under electron microscopy. Furthermore, the recombinant N-terminal domain of MLKL forms amyloid-like fibers and binds Congo red dye. MLKL mutants that cannot form polymers also fail to induce necroptosis efficiently. Finally, the compound necrosulfonamide conjugates cysteine 86 of human MLKL and blocks MLKL polymer formation and subsequent cell death. These results demonstrate that disulfide bond-dependent, amyloid-like MLKL polymers are necessary and sufficient to induce necroptosis.

show abstract

Redox (phospho)lipidomics of signaling in inflammation and programmed cell death

Tyurina

Croix

Watkins

et al. 2019

Journal of Leukocyte Biology

View full text Add to dashboard Cite

In addition to the known prominent role of polyunsaturated (phospho)lipids as structural blocks of biomembranes, there is an emerging understanding of another important function of these molecules as a highly diversified signaling language utilized for intra-and extracellular communications. Technological developments in high-resolution mass spectrometry facilitated the development of a new branch of metabolomics, redox lipidomics. Analysis of lipid peroxidation reactions has already identified specific enzymatic mechanisms responsible for the biosynthesis of several unique signals in response to inflammation and regulated cell death programs. Obtaining comprehensive information about millions of signals encoded by oxidized phospholipids, represented by thousands of interactive reactions and pleiotropic (patho)physiological effects, is a daunting task.However, there is still reasonable hope that significant discoveries, of at least some of the important contributors to the overall overwhelmingly complex network of interactions triggered by inflammation, will lead to the discovery of new small molecule regulators and therapeutic modalities. For example, suppression of the production of AA-derived pro-inflammatory mediators, HXA 3 and LTB 4, by an iPLA 2 inhibitor, R-BEL, mitigated injury associated with the activation of pro-inflammatory processes in animals exposed to whole-body irradiation. Further, technological developments promise to make redox lipidomics a powerful approach in the arsenal of diagnostic and therapeutic instruments for personalized medicine of inflammatory diseases and conditions. K E Y W O R D Seicosanoids, lipid mediators, lipoxygenase, Oxidized phospholipids, peroxidation, phospholipase A2, phospholipid hydrolysis

show abstract

Sequential Engagement of Distinct MLKL Phosphatidylinositol-Binding Sites Executes Necroptosis

Cited by 199 publications

References 28 publications

The MLKL Channel in Necroptosis Is an Octamer Formed by Tetramers in a Dyadic Process

The MLKL Channel in Necroptosis Is an Octamer Formed by Tetramers in a Dyadic Process

MLKL forms disulfide bond-dependent amyloid-like polymers to induce necroptosis

Redox (phospho)lipidomics of signaling in inflammation and programmed cell death

Contact Info

Product

Resources

About