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ZNF451 collaborates with RNF8 to regulate RNF168 localization and amplify ubiquitination signaling to promote DNA damage repair and regulate radiosensitivity

Cell Death & Differentiation volume 32, pages 1303–1316 (2025)Cite this article

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Abstract

The ubiquitination of histone H2A/H2AX, catalyzed by RNF8/RNF168, is a crucial step in the repair of DNA double-strand breaks (DSBs), playing a significant role in transmitting and amplifying DNA damage response signals. However, the upstream regulatory mechanisms of RNF168 remain unclear. Here, we demonstrate that ZNF451 catalyzes the SUMOylation of RNF168, thereby regulating the ubiquitination of histone H2A/H2AX. Specifically, ZNF451 rapidly responds to radiation-induced DNA damage, accumulating abundantly at damage sites and catalyzing the SUMO2 modification of RNF168. This modification stabilizes RNF168, enhancing its accumulation at damage sites, which increases the ubiquitination levels of downstream histone H2A/H2AX and promotes the DNA damage repair process. Furthermore, we find that ZNF451 and RNF8 jointly regulate RNF168 in a novel manner, exhibiting both competitive and cooperative characteristics. The interaction between RNF168 and either ZNF451 or RNF8 mutually inhibits each other. However, simultaneous loss of ZNF451 and RNF8 markedly impedes the recruitment of RNF168 to damage sites. Whereas, varying expression levels of ZNF451 and RNF8 suggest that both facilitate the interaction between RNF168 and the downstream factor H2AX, but the interaction plateaus beyond a specific threshold. Altogether, these findings reveal that the SUMOylation catalyzed by ZNF451 is involved in regulating RNF168-induced ubiquitin signaling in DSBs repair and suggest that ZNF451 could serve as a potential therapeutic target in tumor radiotherapy.

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Fig. 1: ZNF451 is required for DNA damage repair.
Fig. 2: The absence of ZNF451 impairs DDR and enhances radio/chemo-sensitivity of tumor cells.
Fig. 3: ZNF451 regulates non-homologous end joining and homologous recombination repair of DSBs.
Fig. 4: ZNF451 interacts with RNF168 and stabilizes its protein.
Fig. 5: ZNF451 catalyzes the SUMOylation of RNF168 and amplifies the ubiquitin signaling in DNA repair pathway.
Fig. 6: ZNF451 and RNF8 cooperatively regulate RNF168-induced ubiquitin signaling.
Fig. 7: Targeting ZNF451 enhances the efficacy of radiotherapy in vivo.
Fig. 8: Schematic illustration of ZNF451 regulating DNA damage repair mechanism.

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Data availability

Other data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the National Science Fund for Excellent Young Scholars (12122510), the National Natural Science Foundation of China (32171240, 12435019, 32401029 and 32271283), the HFIPS Director’s Fund (BJPY2021B07 and BJPY2023A010).

Author information

Authors and Affiliations

  1. High Magnetic Field Laboratory, Key Laboratory of High Magnetic Field and Ion Beam Physical Biology, Chinese Academy of Sciences; Anhui Province Key Laboratory of Environmental Toxicology and Pollution Control Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, China

    Feng Xu, Ruru Wang, Jie Zhang, Xipeng Zhao, Zhaoyang Zhang, Zhicheng Yao, Jie Zhang, Shenglan Zhou, An Xu & Guoping Zhao

  2. University of Science and Technology of China, Hefei, Anhui, 230026, China

    Feng Xu, Qi Xia, Ruru Wang, Jie Zhang, Xipeng Zhao, Zhaoyang Zhang, Zhicheng Yao, Jie Zhang & Shenglan Zhou

  3. Department of Oncology, Mayo Clinic, Rochester, MN, 55905, USA

    Bin Chen

  4. Anhui Medical University, Hefei, Anhui, 230032, China

    Xiaona Li

  5. Information Materials and Intelligent Sensing Laboratory of Anhui Province, Institutes of Physical Science and Information Technology, Anhui University, Hefei, Anhui, 230601, China

    Biao Chen & Lijun Wu

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Contributions

FX, QX, Bin C and GZ designed and conceived of the study. FX, QX, Bin C, RW, JZ, XZ, ZZ, and Biao C conducted experiments. FX, ZY, JZ, SZ, XL, AX and LW analyzed and interpreted the data. FX and GZ wrote the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Guoping Zhao.

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The study is approved by the Ethics Committee of Hefei Institutes of Physical Science, Chinese Academy of Sciences. All methods were performed in accordance with the relevant guidelines and regulations.

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Xu, F., Xia, Q., Chen, B. et al. ZNF451 collaborates with RNF8 to regulate RNF168 localization and amplify ubiquitination signaling to promote DNA damage repair and regulate radiosensitivity. Cell Death Differ 32, 1303–1316 (2025). https://doi.org/10.1038/s41418-025-01472-0

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