ROS-dependent S-palmitoylation activates cleaved and intact gasdermin D

• 成果类型: Article
• 署名作者: Du, Gang; Healy, Liam B.; David, Liron; Walker, Caitlin; El-Baba, Tarick J.; Lutomski, Corinne A.; Goh, Byoungsook; Gu, Bowen; Pi, Xiong; Devant, Pascal; Fontana, Pietro; Dong, Ying; Ma, Xiyu; Miao, Rui; Balasubramanian, Arumugam; Puthenveetil, Robbins; Banerjee, Anirban; Luo, Hongbo R.; Kagan, Jonathan C.; Oh, Sungwhan F.; Robinson, Carol V.; Lieberman, Judy; Wu, Hao
• 署名单位: Harvard University; Harvard Medical School; Harvard University; Harvard University Medical Affiliates; Boston Children's Hospital; Program in Cellular & Molecular Medicine (PCMM); University of Oxford; University of Oxford; Harvard University; Harvard University Medical Affiliates; Brigham & Women's Hospital; Harvard University; Harvard Medical School; Harvard University; Harvard Medical School; Harvard University; Harvard University Medical Affiliates; Boston Children's Hospital; Harvard University; Harvard Medical School; Harvard University Medical Affiliates; Dana-Farber Cancer Institute; Harvard University; Harvard University Medical Affiliates; Boston Children's Hospital; National Institutes of Health (NIH) - USA; NIH Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD); Ben-Gurion University of the Negev; CSL
• 刊物名称: Nature
• ISSN/ISSBN: 0028-3844
• DOI: 10.1038/s41586-024-07373-5
• 发表日期: 2024-06-13
• 关键词: mass-spectrometry; cell-death; pyroptosis; Visualization; caspase-11; mechanism; gsdmd

摘要

Gasdermin D (GSDMD) is the common effector for cytokine secretion and pyroptosis downstream of inflammasome activation and was previously shown to form large transmembrane pores after cleavage by inflammatory caspases to generate the GSDMD N-terminal domain (GSDMD-NT)1-10. Here we report that GSDMD Cys191 is S-palmitoylated and that palmitoylation is required for pore formation. S-palmitoylation, which does not affect GSDMD cleavage, is augmented by mitochondria-generated reactive oxygen species (ROS). Cleavage-deficient GSDMD (D275A) is also palmitoylated after inflammasome stimulation or treatment with ROS activators and causes pyroptosis, although less efficiently than palmitoylated GSDMD-NT. Palmitoylated, but not unpalmitoylated, full-length GSDMD induces liposome leakage and forms a pore similar in structure to GSDMD-NT pores shown by cryogenic electron microscopy. ZDHHC5 and ZDHHC9 are the major palmitoyltransferases that mediate GSDMD palmitoylation, and their expression is upregulated by inflammasome activation and ROS. The other human gasdermins are also palmitoylated at their N termini. These data challenge the concept that cleavage is the only trigger for GSDMD activation. They suggest that reversible palmitoylation is a checkpoint for pore formation by both GSDMD-NT and intact GSDMD that functions as a general switch for the activation of this pore-forming family.