Dna damage-inducible transcript 3 positively regulates ripk1-mediated necroptosis

ABSTRACT DNA damage-inducible transcript 3 (DDIT3) is a well-known transcription factor that regulates the expression of apoptosis-related genes for promoting apoptosis during endoplasmic

reticulum stress. Here, we report an unrecognized role of DDIT3 in facilitating necroptosis. DDIT3 directly binds and competitively prevents the p38 MAPK-MK2 interaction and thereby blocking

MK2 activation while stimulating p38 MAPK activation. This blockage of MK2 activation initially prevents RIPK1 phosphorylation at Ser320 (inactivation), subsequently relieving its

suppression of RIPK1 activation. Consequently, p38 MAPK facilitates RIPK1 phosphorylation at Ser166 (activation) through DDIT3 phosphorylation-related mechanisms, leading to necroptosis.

Mechanistically, a 10-amino acid segment (Glu19-Val28) within DDIT3’s N-terminus is identified to account for its pro-necroptotic function. In vivo studies demonstrate that forced expression

of DDIT3 induces necroptosis, whereas deletion of DDIT3 alleviates necroptosis in mouse hearts under stress. These findings shed light on a novel regulatory mechanism by which DDIT3

promotes RIPK1 activation and subsequent necroptosis. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS

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STABILIZING PARP1-DNA CONDENSATES Article 27 November 2024 P53 INDUCES ARTS TO PROMOTE MITOCHONDRIAL APOPTOSIS Article Open access 24 February 2021 IKK-MEDIATED TRAF6 AND RIPK1 INTERACTION

STIFLES CELL DEATH COMPLEX ASSEMBLY LEADING TO THE SUPPRESSION OF TNF-Α-INDUCED CELL DEATH Article 21 April 2023 DATA AVAILABILITY The datasets generated during and/or analyzed during the

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Scholar  Download references FUNDING This study was partially supported by a discovery grant from the Natural Sciences and Engineering Research Council of Canada (RGPIN-2023-04573 to TP),

the Lawson Internal Research Fund (TP), High-level Foreign Experts Project by the Ministry of Science and Technology (G2022014014156L), Research Cooperation and High-level Talent Training

Project of the China Scholarship Council (2022), and Projects of International Cooperation from Jiangsu (BZ2024031, China). AUTHOR INFORMATION Author notes * These authors contributed

equally: Rui Ni, Ting Cao. AUTHORS AND AFFILIATIONS * International Genome Center, Jiangsu University, Zhenjiang, 212013, China Rui Ni, Xiaoyun Ji & Zhaoliang Su * Lawson Health Research

Institute, London Health Sciences Centre, London, ON, N6A 5W9, Canada Rui Ni, Xiaoyun Ji, Zhuxu Zhang & Tianqing Peng * Department of Pathology and Laboratory Medicine, Western

University, London, ON, N6A 5C1, Canada Rui Ni, Xiaoyun Ji, Angel Peng, Zhuxu Zhang, Subrata Chakrabarti & Tianqing Peng * Institute for Cardiovascular Science, Soochow University,

Suzhou, 215123, China Ting Cao * Department of Medicine, Western University, London, ON, N6A 5W9, Canada Zhuxu Zhang & Tianqing Peng * Department of Pharmacology and Systems Physiology,

University of Cincinnati College of Medicine, Cincinnati, OH, 45267, USA Guo-Chang Fan * Department of Physiology and Pharmacology, Western University, London, ON, N6A 5C1, Canada Peter

Stathopulos Authors * Rui Ni View author publications You can also search for this author inPubMed Google Scholar * Ting Cao View author publications You can also search for this author

inPubMed Google Scholar * Xiaoyun Ji View author publications You can also search for this author inPubMed Google Scholar * Angel Peng View author publications You can also search for this

author inPubMed Google Scholar * Zhuxu Zhang View author publications You can also search for this author inPubMed Google Scholar * Guo-Chang Fan View author publications You can also search

for this author inPubMed Google Scholar * Peter Stathopulos View author publications You can also search for this author inPubMed Google Scholar * Subrata Chakrabarti View author

publications You can also search for this author inPubMed Google Scholar * Zhaoliang Su View author publications You can also search for this author inPubMed Google Scholar * Tianqing Peng

View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS RN, TC, XJ, and AP conducted the experiments and analyzed the data; RN, TC, GCF, ZZ, SC,

ZS, and TP designed the experiments and wrote the paper; RN, XJ, AP, GCF, ZZ, PS, SC, ZS, and TP discussed the data and revised the paper. PS contributed to protein-protein docking analysis

and structural mechanism. All authors approved the final version to be published. CORRESPONDING AUTHORS Correspondence to Zhaoliang Su or Tianqing Peng. ETHICS DECLARATIONS COMPETING

INTERESTS The authors declare no competing interests. ETHICAL APPROVAL All mouse experimental procedures were approved by the Animal Use Subcommittee at the University of Western Ontario,

Canada (protocol numbers: 2021-054 and 2020-047), and Soochow University, China (protocol number: 2017-0043). All methods were performed in accordance with the relevant guidelines and

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applicable law. Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Ni, R., Cao, T., Ji, X. _et al._ DNA damage-inducible transcript 3 positively regulates RIPK1-mediated

necroptosis. _Cell Death Differ_ 32, 306–319 (2025). https://doi.org/10.1038/s41418-024-01385-4 Download citation * Received: 26 December 2023 * Revised: 12 September 2024 * Accepted: 17

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