Seo, Jinho Lee, Eun-Woo Shin, Jihye Seong, Daehyeon Nam, Young Woo Jeong, Manhyung Lee, Seon-Hyeong Lee, Cheolju Song, Jaewhan 2024-01-19T22:00:37Z 2024-01-19T22:00:37Z 2021-09-03 2018-09-06 0950-9232 https://pubs.kist.re.kr/handle/201004/120922 Fas-associated death domain (FADD) is an adaptor protein recruiting complexes of caspase 8 to death ligand receptors to induce extrinsic apoptotic cell death in response to a TNF superfamily member. Although, formation of the complex of FADD and caspase 8 upon death stimuli has been studied in detail, posttranslational modifications fine-tuning these processes have yet to be identified. Here we revealed that K6-linked polyubiquitylation of FADD on lysines 149 and 153 mediated by C terminus HSC70-interacting protein (CHIP) plays an important role in preventing formation of the death inducing signaling complex (DISC), thus leading to the suppression of cell death. Cells depleted of CHIP showed higher sensitivity toward death ligands such as FasL and TRAIL, leading to upregulation of DISC formation composed of a death receptor, FADD, and caspase 8. CHIP was able to bind to FADD, induce K6-linked polyubiquitylation of FADD, and suppress DISC formation. By mass spectrometry, lysines 149 and 153 of FADD were found to be responsible for CHIP-mediated FADD ubiquitylation. FADD mutated at these sites was capable of more potent cell death induction as compared with the wild type and was no longer suppressed by CHIP. On the other hand, CHIP deficient in E3 ligase activity was not capable of suppressing FADD function and of FADD ubiquitylation. CHIP depletion in ME-180 cells induced significant sensitization of these cells toward TRAIL in xenograft analyses. These results imply that K6-linked ubiquitylation of FADD by CHIP is a crucial checkpoint in cytokine-dependent extrinsic apoptosis. English NATURE PUBLISHING GROUP UBIQUITIN LIGASE CHIP CONTAINING PROTEIN APOPTOSIS FAS PHOSPHORYLATION KINASE DOMAIN CASPASE-8 RECEPTOR CHAINS K6 linked polyubiquitylation of FADD by CHIP prevents death inducing signaling complex formation suppressing cell death Article 10.1038/s41388-018-0323-z 1 ONCOGENE, v.37, no.36, pp.4994 - 5006 ONCOGENE 37 36 4994 5006 scie scopus 000443823600007 2-s2.0-85047263072 Biochemistry & Molecular Biology Oncology Cell Biology Genetics & Heredity Biochemistry & Molecular Biology Oncology Cell Biology Genetics & Heredity Article UBIQUITIN LIGASE CHIP CONTAINING PROTEIN APOPTOSIS FAS PHOSPHORYLATION KINASE DOMAIN CASPASE-8 RECEPTOR CHAINS