Z-REX uncovers a bifurcation in function of Keap1 paralogs
Studying electrophile signaling is marred by difficulties in parsing changes in pathway flux attributable to on-target, vis-à-vis off-target, modifications. By combining bolus dosing, knockdown, and Z-REX—a tool investigating on-target/on-pathway electrophile signaling, we document that electrophile...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-10-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/83373 |
| _version_ | 1827935908721065984 |
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| author | Alexandra Van Hall-Beauvais Jesse R Poganik Kuan-Ting Huang Saba Parvez Yi Zhao Hong-Yu Lin Xuyu Liu Marcus John Curtis Long Yimon Aye |
| author_facet | Alexandra Van Hall-Beauvais Jesse R Poganik Kuan-Ting Huang Saba Parvez Yi Zhao Hong-Yu Lin Xuyu Liu Marcus John Curtis Long Yimon Aye |
| author_sort | Alexandra Van Hall-Beauvais |
| collection | DOAJ |
| description | Studying electrophile signaling is marred by difficulties in parsing changes in pathway flux attributable to on-target, vis-à-vis off-target, modifications. By combining bolus dosing, knockdown, and Z-REX—a tool investigating on-target/on-pathway electrophile signaling, we document that electrophile labeling of one zebrafish-Keap1-paralog (zKeap1b) stimulates Nrf2- driven antioxidant response (AR) signaling (like the human-ortholog). Conversely, zKeap1a is a dominant-negative regulator of electrophile-promoted Nrf2-signaling, and itself is nonpermissive for electrophile-induced Nrf2-upregulation. This behavior is recapitulated in human cells: (1) zKeap1b-expressing cells are permissive for augmented AR-signaling through reduced zKeap1b–Nrf2 binding following whole-cell electrophile treatment; (2) zKeap1a-expressing cells are non-permissive for AR-upregulation, as zKeap1a–Nrf2 binding capacity remains unaltered upon whole-cell electrophile exposure; (3) 1:1 ZKeap1a:zKeap1b-co-expressing cells show no Nrf2-release from the Keap1-complex following whole-cell electrophile administration, rendering these cells unable to upregulate AR. We identified a zKeap1a-specific point-mutation (C273I) responsible for zKeap1a’s behavior during electrophilic stress. Human-Keap1(C273I), of known diminished Nrf2-regulatory capacity, dominantly muted electrophile-induced Nrf2-signaling. These studies highlight divergent and interdependent electrophile signaling behaviors, despite conserved electrophile sensing. |
| first_indexed | 2024-03-13T08:01:40Z |
| format | Article |
| id | doaj.art-eda320ac548e43cfb8ab7eb2372f9c20 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-03-13T08:01:40Z |
| publishDate | 2022-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-eda320ac548e43cfb8ab7eb2372f9c202023-06-01T14:06:47ZengeLife Sciences Publications LtdeLife2050-084X2022-10-011110.7554/eLife.83373Z-REX uncovers a bifurcation in function of Keap1 paralogsAlexandra Van Hall-Beauvais0https://orcid.org/0000-0003-2515-5191Jesse R Poganik1Kuan-Ting Huang2https://orcid.org/0000-0001-7057-1448Saba Parvez3Yi Zhao4https://orcid.org/0000-0002-6049-1943Hong-Yu Lin5Xuyu Liu6Marcus John Curtis Long7Yimon Aye8https://orcid.org/0000-0002-1256-4159Swiss Federal Institute of Technology Lausanne, Lausanne, SwitzerlandSwiss Federal Institute of Technology Lausanne, Lausanne, Switzerland; Division of Genetics, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, United StatesSwiss Federal Institute of Technology Lausanne, Lausanne, SwitzerlandDepartment of Pharmacology and Toxicology, College of Pharmacy, University of Utah, Salt Lake City, United StatesSwiss Federal Institute of Technology Lausanne, Lausanne, Switzerland; BayRay Innovation Center, Shenzhen Bay Laboratory, Shenzhen, ChinaDepartment of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, ChinaSwiss Federal Institute of Technology Lausanne, Lausanne, Switzerland; School of Chemistry, The University of Sydney, Sydney, Australia; The Heart Research Institute, Newtown, Newtown, AustraliaDepartment of Biochemistry, Faculty of Biology and Medicine, University of Lausanne, Lausanne, SwitzerlandSwiss Federal Institute of Technology Lausanne, Lausanne, SwitzerlandStudying electrophile signaling is marred by difficulties in parsing changes in pathway flux attributable to on-target, vis-à-vis off-target, modifications. By combining bolus dosing, knockdown, and Z-REX—a tool investigating on-target/on-pathway electrophile signaling, we document that electrophile labeling of one zebrafish-Keap1-paralog (zKeap1b) stimulates Nrf2- driven antioxidant response (AR) signaling (like the human-ortholog). Conversely, zKeap1a is a dominant-negative regulator of electrophile-promoted Nrf2-signaling, and itself is nonpermissive for electrophile-induced Nrf2-upregulation. This behavior is recapitulated in human cells: (1) zKeap1b-expressing cells are permissive for augmented AR-signaling through reduced zKeap1b–Nrf2 binding following whole-cell electrophile treatment; (2) zKeap1a-expressing cells are non-permissive for AR-upregulation, as zKeap1a–Nrf2 binding capacity remains unaltered upon whole-cell electrophile exposure; (3) 1:1 ZKeap1a:zKeap1b-co-expressing cells show no Nrf2-release from the Keap1-complex following whole-cell electrophile administration, rendering these cells unable to upregulate AR. We identified a zKeap1a-specific point-mutation (C273I) responsible for zKeap1a’s behavior during electrophilic stress. Human-Keap1(C273I), of known diminished Nrf2-regulatory capacity, dominantly muted electrophile-induced Nrf2-signaling. These studies highlight divergent and interdependent electrophile signaling behaviors, despite conserved electrophile sensing.https://elifesciences.org/articles/83373Keap1/Nrf2 antioxidant responseelectrophile signalingparalog-specific signalingzebrafishREX technologiescovalent drug mode-of-action |
| spellingShingle | Alexandra Van Hall-Beauvais Jesse R Poganik Kuan-Ting Huang Saba Parvez Yi Zhao Hong-Yu Lin Xuyu Liu Marcus John Curtis Long Yimon Aye Z-REX uncovers a bifurcation in function of Keap1 paralogs eLife Keap1/Nrf2 antioxidant response electrophile signaling paralog-specific signaling zebrafish REX technologies covalent drug mode-of-action |
| title | Z-REX uncovers a bifurcation in function of Keap1 paralogs |
| title_full | Z-REX uncovers a bifurcation in function of Keap1 paralogs |
| title_fullStr | Z-REX uncovers a bifurcation in function of Keap1 paralogs |
| title_full_unstemmed | Z-REX uncovers a bifurcation in function of Keap1 paralogs |
| title_short | Z-REX uncovers a bifurcation in function of Keap1 paralogs |
| title_sort | z rex uncovers a bifurcation in function of keap1 paralogs |
| topic | Keap1/Nrf2 antioxidant response electrophile signaling paralog-specific signaling zebrafish REX technologies covalent drug mode-of-action |
| url | https://elifesciences.org/articles/83373 |
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