Mingtong Ma, Fei Wang, Peng‐Fei Cui, Yingying Zhang, Hanyu Ma, Yifan He, Yuanna Cheng, Jingping Huang, Jingxiang Wang, Xiangyang Wu, Hua Yang, Ruijuan Zheng, Haowen Ma, Qian Cai, Lin Wang, Baoxue Ge
Macrophage infection by the pathogenic bacteria Yersinia or mimic stimulation of lipopolysaccharide (LPS) and transforming growth factor-β-activated kinase 1 (TAK1) inhibitor or tumor necrosis factor (TNF) and TAK1 inhibitor induces caspase-8-mediated gasdermin D (GSDMD) cleavage and pyroptosis. However, the upstream regulator of caspase-8-dependent cleavage of GSDMD remains elusive. Here we show that Serine/threonine-protein kinase RIO2 (RIOK2) interacts with the Fas-associated protein with death domain (FADD) and is essential for caspase-8-driven GSDMD cleavage. RIOK2’s kinase activity drives the transport of lysosome to ER through activating myosin II and thereby translocate FADD-RIPK1-caspase-8 complex from lysosome to ER. Importantly, RIOK2’s ATPase activity enhances its binding to this complex and directly triggers caspase-8 and gasdermin D cleavage both at ER and in vitro. Furthermore, RIOK2-mediated pyroptosis enhances host defense against Yersinia infection. Thus, our findings define an upstream regulator of caspase-8-dependent pyroptosis, implying a role of organelle crosstalk in spatial cleavage of gasdermins. Macrophage infection by Yersinia or specific stimulations trigger caspase-8-mediated GSDMD cleavage. RIOK2 kinase is essential for this process by interacting with FADD and driving the lysosome-to-ER transport of the cell death complex accompanied by GSDMD cleavage, thereby enhancing host defense.