Homo sapiens Raw sequence reads. Homo sapiens

Gasdermin D is the common effector for cytokine secretion and pyroptosis downstream of inflammasome activation and was previously shown to form large transmembrane pores upon cleavage by inflammatory caspases. Here we report that S palmitoylation at Cys191 of GSDMD enables pore formation by both cleaved GSDMD N terminal domain and intact GSDMD, leading to cell death and IL1B release. Palmitoylation is induced by inflammasome activation shown for multiple inflammasomes and stimuli, suppressed by the palmitoylation inhibitor 2 bromopalmitate and the C191A mutation without affecting GSDMD cleavage, and augmented and abolished, respectively, by activators and quenchers of reactive oxygen species. Inflammasome activation induced GSDMD palmitoylation is dependent on a feed forward loop, as palmitoylated GSDMD damages mitochondria, leading to more ROS, which in turn enhances palmitoylation. zDHHC5 and zDHHC9 are the major palmitoyl transferases for mediating GSDMD palmitoylation, and their expression is upregulated by inflammasome activation and ROS through reducing proteasomal degradation. RNAseq data was used to analyze the expression level after the treatment of 2BP or NAC.