Consecutive palmitoylation and phosphorylation orchestrates NLRP3 membrane trafficking and inflammasome activation

• Published in: Molecular cell Vol. 84; no. 17; pp. 3336 - 3353.e7
• Main Authors: Nie, Li, Fei, Chenjie, Fan, Yizeng, Dang, Fabin, Zhao, Ziyue, Zhu, Tingfang, Wu, Xiangyu, Dai, Ting, Balasubramanian, Arumugam, Pan, Jing, Hu, Yang, Luo, Hongbo R., Wei, Wenyi, Chen, Jiong
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.09.2024
• Subjects: Acyltransferases - genetics; Acyltransferases - metabolism; Animals; Endosomes - metabolism; HEK293 Cells; Humans; inflammasome activation; Inflammasomes - genetics; Inflammasomes - metabolism; LATS1/2; Lipoylation; membrane trafficking; Mice; Mice, Inbred C57BL; Mice, Knockout; microtubule-organizing center; Microtubule-Organizing Center - metabolism; Mitochondria - metabolism; NIMA-Related Kinases - genetics; NIMA-Related Kinases - metabolism; NLR Family, Pyrin Domain-Containing 3 Protein - genetics; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; NLRP3; palmitoylation; Phosphorylation; Protein Transport; trans-Golgi Network - metabolism; zDHHC1; inflammasome activation; microtubule-organizing center; zDHHC1; NLRP3; LATS1/2; phosphorylation; palmitoylation; membrane trafficking
• ISSN: 1097-2765; 1097-4164; 1097-4164
• DOI: 10.1016/j.molcel.2024.08.001

NLRP3 inflammasome activation, essential for cytokine secretion and pyroptosis in response to diverse stimuli, is closely associated with various diseases. Upon stimulation, NLRP3 undergoes subcellular membrane trafficking and conformational rearrangements, preparing itself for inflammasome assembly at the microtubule-organizing center (MTOC). Here, we elucidate an orchestrated mechanism underlying these ordered processes using human and murine cells. Specifically, NLRP3 undergoes palmitoylation at two sites by palmitoyl transferase zDHHC1, facilitating its trafficking between subcellular membranes, including the mitochondria, trans-Golgi network (TGN), and endosome. This dynamic trafficking culminates in the localization of NLRP3 to the MTOC, where LATS1/2, pre-recruited to MTOC during priming, phosphorylates NLRP3 to further facilitate its interaction with NIMA-related kinase 7 (NEK7), ultimately leading to full NLRP3 activation. Consistently, Zdhhc1-deficiency mitigated LPS-induced inflammation and conferred protection against mortality in mice. Altogether, our findings provide valuable insights into the regulation of NLRP3 membrane trafficking and inflammasome activation, governed by palmitoylation and phosphorylation events. [Display omitted] •Palmitoylation of NLRP3 at C130 and C958 is critical for its membrane trafficking•zDHHC1 serves as the primary palmitoyl transferase for NLRP3 at C130 and C958•LATS1/2 phosphorylates NLRP3 at S265 to promote its interaction with NEK7 at the MTOC•zDHHC1-deficiency mitigates LPS-induced inflammation and enhances survival in mice Nie et al. discovered that zDHHC1-mediated palmitoylation of NLRP3 at C130 and C958 residues directs its membrane localization and trafficking to the MTOC, where LATS1/2-mediated phosphorylation facilitates NEK7 interaction and inflammasome assembly. This sequential regulation is crucial for NLRP3 activation, highlighting potential therapeutic targets for controlling inflammation.