title: Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death.
abstract: Inflammatory caspases (caspase-1, -4, -5 and -11) are critical for innate defences. Caspase-1 is activated by ligands of various canonical inflammasomes, and caspase-4, -5 and -11 directly recognize bacterial lipopolysaccharide, both of which trigger pyroptosis. Despite the crucial role in immunity and endotoxic shock, the mechanism for pyroptosis induction by inflammatory caspases is unknown. Here we identify gasdermin D (Gsdmd) by genome-wide clustered regularly interspaced palindromic repeat (CRISPR)-Cas9 nuclease screens of caspase-11- and caspase-1-mediated pyroptosis in mouse bone marrow macrophages. GSDMD-deficient cells resisted the induction of pyroptosis by cytosolic lipopolysaccharide and known canonical inflammasome ligands. Interleukin-1β release was also diminished in Gsdmd(-/-) cells, despite intact processing by caspase-1. Caspase-1 and caspase-4/5/11 specifically cleaved the linker between the amino-terminal gasdermin-N and carboxy-terminal gasdermin-C domains in GSDMD, which was required and sufficient for pyroptosis. The cleavage released the intramolecular inhibition on the gasdermin-N domain that showed intrinsic pyroptosis-inducing activity. Other gasdermin family members were not cleaved by inflammatory caspases but shared the autoinhibition; gain-of-function mutations in Gsdma3 that cause alopecia and skin defects disrupted the autoinhibition, allowing its gasdermin-N domain to trigger pyroptosis. These findings offer insight into inflammasome-mediated immunity/diseases and also change our understanding of pyroptosis and programmed necrosis.
authors.affiliation: Peking University-Tsinghua University-National Institute of Biological Sciences Joint Graduate Program, School of Life Sciences, Tsinghua University, 100084, China.|National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Jianjin
authors.initials: J
authors.lastname: Shi
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Xuyan
authors.initials: X
authors.lastname: Shi
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Huanwei
authors.initials: H
authors.lastname: Huang
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Zhuang
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Cai
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Fengchao
authors.initials: F
authors.lastname: Wang
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.|National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.|National Institute of Biological Sciences, Beijing, Collaborative Innovation Center for Cancer Medicine, Beijing 102206, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Shao
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D064113:CRISPR-Cas Systems; D020170:Caspase 1 / Q000378:metabolism; D020169:Caspases / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism; D007249:Inflammation / Q000201:enzymology*; D053583:Interleukin-1beta / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009336:Necrosis; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism*; D028044:Phosphate-Binding Proteins; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D000094524:Gasdermins
chemical_list: D051017:Apoptosis Regulatory Proteins; D020170:Caspase 1; D020169:Caspases; D058847:Inflammasomes; D053583:Interleukin-1beta; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D009363:Neoplasm Proteins; D028044:Phosphate-Binding Proteins; D011506:Proteins; C498366:GSDMD protein, human; D000094524:Gasdermins; C499121:Gsdma3 protein, mouse; C498368:Gsdmd protein, mouse