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Title:
Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death | Nature
Description:
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. CRISPR-Cas9 genome-editing screens identify gasdermin D as a substrate for inflammatory caspases, and its N-terminal cleavage fragment, as well as the equivalent regions in other gasdermins, is shown to be capable of inducing pyroptosis. Two groups reporting in this issue of Nature identify gasdermin D, the product of the Gsdmd gene conserved in human and mouse but of unknown function, as a substrate for inflammatory caspases. Feng Shao and co-workers use genome-wide CRISPR–Cas9 screens to identify gasdermin D as a substrate for inflammatory caspases. Caspase-1 and caspase-4/5/11 specifically cleaved the linker between the amino-terminal gasdermin-N and carboxy-terminal gasdermin-C domains. Vishva Dixit and co-workers use an ENU mutagenesis screen to identify gasdermin D as the required substrate for pyroptosis-mediating caspase-11 in the non-canonical inflammasome pathway. Mice lacking gasdermin D are protected from a lethal dose of lipopolysaccharide. Both groups show that the cleaved N-terminal domain is sufficient to trigger pyroptosis, a form of programmed necrotic cell death.
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nature portfolio permissions reprints privacy policy advertising genome-wide crispr–cas9 screens genome-wide crispr-cas9 screen social media nature identify gasdermin acute lung injury fr/espript/cgi-bin/espript atp-based cell viability research nlrp1-dependent pyroptosis leads lfn–bsak-induced gsdmd cleavage crispr-cas9-mediated targeting del castillo velasco-herrera nature biotechnol crispr-cas9-mdiated targeting lfn–bsak-triggered pyroptosis nature immunol reverse-transcription pcr analyses inflammasome-mediated caspase-1 autoprocessing lfn–bsak-induced pyroptosis prolonged lfn–bsak treatment overexpression-activated inflammatory caspases inflammasome-mediated immunity/diseases pan-caspase inhibitor zvad author correspondence bacterial-infection-activated caspase-1 macrophages drives orchitis gsdmd-deficient cell lines nature 477 nature 513 nature 514 nature 479 nature 505 nature 490 nature 526 nature t3ss-deficient mutant strains lps electroporation-induced pyroptosis caspase-1-mediated caspase-3/7 cleavage t3ss-deficient δsipd mutant boosts anti-tumor immunity gsdmd-deficient cells resisted interleukin-1β release springerlink instant access anti-tubulin blots serving tlr4-independent endotoxic shock n-terminal cleavage product
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headline:Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death
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description: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 Gsdmdâ/â 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. CRISPR-Cas9 genome-editing screens identify gasdermin D as a substrate for inflammatory caspases, and its N-terminal cleavage fragment, as well as the equivalent regions in other gasdermins, is shown to be capable of inducing pyroptosis. Two groups reporting in this issue of Nature identify gasdermin D, the product of the Gsdmd gene conserved in human and mouse but of unknown function, as a substrate for inflammatory caspases. Feng Shao and co-workers use genome-wide CRISPRâCas9 screens to identify gasdermin D as a substrate for inflammatory caspases. Caspase-1 and caspase-4/5/11 specifically cleaved the linker between the amino-terminal gasdermin-N and carboxy-terminal gasdermin-C domains. Vishva Dixit and co-workers use an ENU mutagenesis screen to identify gasdermin D as the required substrate for pyroptosis-mediating caspase-11 in the non-canonical inflammasome pathway. Mice lacking gasdermin D are protected from a lethal dose of lipopolysaccharide. Both groups show that the cleaved N-terminal domain is sufficient to trigger pyroptosis, a form of programmed necrotic cell death.
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