title: Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase.
abstract: The receptor-interacting serine-threonine kinase 3 (RIP3) is a key signaling molecule in the programmed necrosis (necroptosis) pathway. This pathway plays important roles in a variety of physiological and pathological conditions, including development, tissue damage response, and antiviral immunity. Here, we report the identification of a small molecule called (E)-N-(4-(N-(3-methoxypyrazin-2-yl)sulfamoyl)phenyl)-3-(5-nitrothiophene-2-yl)acrylamide--hereafter referred to as necrosulfonamide--that specifically blocks necrosis downstream of RIP3 activation. An affinity probe derived from necrosulfonamide and coimmunoprecipitation using anti-RIP3 antibodies both identified the mixed lineage kinase domain-like protein (MLKL) as the interacting target. MLKL was phosphorylated by RIP3 at the threonine 357 and serine 358 residues, and these phosphorylation events were critical for necrosis. Treating cells with necrosulfonamide or knocking down MLKL expression arrested necrosis at a specific step at which RIP3 formed discrete punctae in cells. These findings implicate MLKL as a key mediator of necrosis signaling downstream of the kinase RIP3.
authors.affiliation: National Institute of Biological Sciences, 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.
authors.forename: Liming
authors.initials: L
authors.lastname: Sun
authors.forename: Huayi
authors.initials: H
authors.lastname: Wang
authors.forename: Zhigao
authors.initials: Z
authors.lastname: Wang
authors.forename: Sudan
authors.initials: S
authors.lastname: He
authors.forename: She
authors.initials: S
authors.lastname: Chen
authors.forename: Daohong
authors.initials: D
authors.lastname: Liao
authors.forename: Lai
authors.initials: L
authors.lastname: Wang
authors.forename: Jiacong
authors.initials: J
authors.lastname: Yan
authors.forename: Weilong
authors.initials: W
authors.lastname: Liu
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Lei
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Wang
mesh_terms: D000178:Acrylamides / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D055785:Gene Knockdown Techniques; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009336:Necrosis / Q000378:metabolism*; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000378:metabolism*; D016415:Sequence Alignment; D015398:Signal Transduction; D013449:Sulfonamides / Q000494:pharmacology
chemical_list: D000178:Acrylamides; C570695:N-(4-(N-(3-methoxypyrazin-2-yl)sulfamoyl)phenyl)-3-(5-nitrothiophene-2-yl)acrylamide; D013449:Sulfonamides; C570693:MLKL protein, human; C570694:MLKL protein, mouse; D011494:Protein Kinases; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C485615:Ripk3 protein, mouse