UNRAVELING THE DUB NETWORK THAT CONTROLS DEATH RECEPTOR INDUCED CELL SURVIVAL AND DEATH PATHWAYS

• Applicant: Privatdozent Dr. Sjoerd van Wijk
• Subject Area: Cell Biology ; Biochemistry
• Term: from 2018 to 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 414826869

• Final Report Year: 2022

Final Report Abstract

Appropriate control of programmed cell death (e.g. apoptosis and necroptosis) and survival underlies embryonal development, innate immunity and cancer surveillance. Disturbances in cell fate checkpoints affect the inflammatory state of the cellular microenvironment, but the molecular mechanisms of how cells decide and switch between death/survival pathways remain poorly understood. Tumor necrosis factor (TNF) receptor 1 (TNFR1) is a prototypical death receptor and a central node in controlling apoptotic, necroptotic and survival responses. Post-translational modification of downstream TNFR1 substrate proteins with linear (M1) and K63-linked ubiquitin chains is essential for controlling these checkpoints and survival/death switching. M1-linked ubiquitination is catalyzed by the LUBAC E3 ligase and counteracted by deubiquitinating enzymes (DUBs), such as OTULIN. However, the interplay of E3-DUB and DUB-DUB pairs for the regulation of ubiquitin-dependent cell fate checkpoints and switching remains largely unclear. Furthermore, numerous key proteins involved in cell death/survival are ubiquitinated as well, but regulatory DUBs for these crucial ubiquitination events remain unidentified. Therefore, a systemic approach to understand the complex E3-DUB and DUB-DUB interplay will reveal novel insights into the vital control of cell survival/death responses. Overall, this proposal aimed to unravel the role of specific ubiquitin signals in controlling TNFR1-mediated survival and programmed cell death pathways in mammalian cells and to investigate the contribution of additional DUBs to cell fate switching. We identified novel signaling functions of LUBAC-mediated M1 ubiquitination during necroptosis, developed a single-molecule resolution-based quantitative imaging platform to study the role of ubiquitination during TNFR1-mediated cell fate signaling and identified novel DUBs that control necroptosis and other forms of TNFα-induced programmed cell death. These findings are expected to have important implications for understanding necroptosis and ubiquitin-mediated cell survival signaling in the context of innate immunity, infections and oncogenesis.

Publications

• Differential involvement of TAK1, RIPK1 and NF-κB signaling in Smac mimetic-induced cell death in breast cancer cells. Biological Chemistry, 400(2), 171-180.
  Schmidt, Nadine; Kowald, Lisa; van Wijk Sjoerd, J.L. & Fulda, Simone
  
• Interferons Transcriptionally Up-Regulate MLKL Expression in Cancer Cells. Neoplasia, 21(1), 74-81.
  Knuth, Anne-Kathrin; Rösler, Stefanie; Schenk, Barbara; Kowald, Lisa; van Wijk Sjoerd, J.L. & Fulda, Simone
  
• Visualizing ubiquitination in mammalian cells. EMBO reports, 20(2).
  van Wijk, Sjoerd J.L.; Fulda, Simone; Dikic, Ivan & Heilemann, Mike
  
• Quantitative single-molecule imaging of TNFR1 reveals zafirlukast as antagonist of TNFR1 clustering and TNFα-induced NF-ĸB signaling. Journal of Leukocyte Biology, 109(2), 363-371.
  Weinelt, Nadine; Karathanasis, Christos; Smith, Sonja; Medler, Juliane; Malkusch, Sebastian; Fulda, Simone; Wajant, Harald; Heilemann, Mike & van, Wijk Sjoerd J. L.
  
• Single-molecule imaging reveals the oligomeric state of functional TNFα-induced plasma membrane TNFR1 clusters in cells. Science Signaling, 13(614).
  Karathanasis, Christos; Medler, Juliane; Fricke, Franziska; Smith, Sonja; Malkusch, Sebastian; Widera, Darius; Fulda, Simone; Wajant, Harald; van Wijk Sjoerd, J. L.; Dikic, Ivan & Heilemann, Mike
  
• Ubiquitin-dependent and -independent functions of OTULIN in cell fate control and beyond. Cell Death & Differentiation, 28(2), 493-504.
  Weinelt, Nadine & van Wijk Sjoerd, J. L.
  
• USP22 controls necroptosis by regulating receptor‐interacting protein kinase 3 ubiquitination. EMBO reports, 22(2).
  Roedig, Jens; Kowald, Lisa; Juretschke, Thomas; Karlowitz, Rebekka; Ahangarian, Abhari Behnaz; Roedig, Heiko; Fulda, Simone; Beli, Petra & van, Wijk Sjoerd JL
  
• Smac mimetics and TRAIL cooperate to induce MLKL-dependent necroptosis in Burkitt's lymphoma cell lines. Neoplasia, 23(5), 539-550.
  Koch, Annkathrin; Jeiler, Birte; Roedig, Jens; van Wijk Sjoerd, J.L.; Dolgikh, Nadezda & Fulda, Simone
  
• Surviving death: emerging concepts of RIPK3 and MLKL ubiquitination in the regulation of necroptosis. The FEBS Journal, 290(1), 37-54.
  Karlowitz, Rebekka & van Wijk Sjoerd, J. L.
  
• The Smac mimetic BV6 cooperates with STING to induce necroptosis in apoptosis-resistant pancreatic carcinoma cells. Cell Death & Disease, 12(9).
  Hannes, Sabine; Karlowitz, Rebekka & van Wijk Sjoerd, J. L.