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K6 linked polyubiquitylation of FADD by CHIP prevents death inducing signaling complex formation suppressing cell death

Oncogenevolume 37pages49945006 (2018) | Download Citation

Abstract

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.

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Acknowledgements

This research was supported by a grant from the Ministry of Science, ICT, and Future Planning (2015R1A3A2066581) (Jaewhan S.) and by the Basic Science Research Program through the National Research Foundation of Korea (NRF) Funded by the Ministry of Education (2017R1A6A3A11035262) (Jinho S.). Additionally, this research was partly supported by the BK21 Plus project of the National Research Foundation of Korea Grant (Jinho S., D.S., M.J., and Y.W.N.), and C.L. acknowledges institutional support by KIST.

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Affiliations

  1. Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Korea

    • Jinho Seo
    • , Daehyeon Seong
    • , Young Woo Nam
    • , Manhyung Jeong
    •  & Jaewhan Song
  2. Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Korea

    • Eun-Woo Lee
  3. Center for Theragnosis, Korea Institute of Science and Technology, 5 Hwarang-ro-14-gil, Seoul, 02792, Korea

    • Jihye Shin
    •  & Cheolju Lee
  4. Cancer Cell & Molecular Biology Branch, Division of Cancer Biology, National Cancer Center, Goyang, 10408, Korea

    • Seon-Hyeong Lee

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The authors declare that they have no conflict of interest.

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Correspondence to Jaewhan Song.

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DOI

https://doi.org/10.1038/s41388-018-0323-z