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Phospho-TRIM21 orchestrates RPA2 ubiquitination switch to promote homologous recombination and tumor radio/chemo-resistance

Oncogene volume 44, pages 1106–1117 (2025)Cite this article

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Abstract

RPA2, a key component of the RPA complex, is essential for single-stranded DNA (ssDNA) binding and DNA repair. However, the regulation of RPA2-ssDNA interaction and the recruitment of repair proteins following DNA damage remain incompletely understood. Our study uncovers a novel mechanism by which phosphorylated TRIM21 (Phospho-TRIM21) regulates RPA2 ubiquitination, thereby modulating homologous recombination and tumor radio/chemo-resistance. In the absence of DNA damage, TRIM21 mediates K63-linked ubiquitination of RPA2, countering K6-linked ubiquitination. Upon DNA damage, ubiquitination-modified RPA2 binds ssDNA, stabilizing the DNA structure and facilitating ATRIP/ATR recruitment. ATR subsequently phosphorylates TRIM21 at Ser41, leading to the dissociation of the TRIM21-RPA2 complex and a shift in RPA2 ubiquitination from K63 to K6 linkage. This shift maintains RPA2 ubiquitination homeostasis and stabilizes the RPA2-ATRIP complex, which is crucial for efficient homologous recombination (HR) repair and enhanced tumor radio/chemo-resistance. We also demonstrate that TRIM21 is frequently upregulated in cancers, and its depletion sensitizes cancer cells to radio/chemotherapy, suggesting its potential as a therapeutic target. This study provides novel insights into TRIM21’s role in the DNA damage response and its implications for cancer treatment.

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Fig. 1: Interaction between TRIM21 and RPA2.
Fig. 2: TRIM21 regulates RPA2 ubiquitination: K63 promotion and K6 inhibition in DBD-D.
Fig. 3: TRIM21 depends on its PRYSPRY ___domain for recruitment to DNA damage sites.
Fig. 4: TRIM21 regulates the binding of RPA2 to ssDNA through ubiquitination, promoting the recruitment of ATRIP and driving HR repair.
Fig. 5: ATR-mediated TRIM21 phosphorylation orchestrates RPA2 ubiquitination switch in DDR.
Fig. 6: TRIM21 is implicated in tumor cell survival and carcinogenesis, and its deletion renders cells sensitive to radiation and chemotherapeutic agents.
Fig. 7: Schematic illustration of TRIM21 regulating HR 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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Acknowledgements

This work was supported by the National Science Fund for Excellent Young Scholars (12122510), the National Natural Science Foundation of China (32171240), the HFIPS Director’s Fund (BJPY2021B07 and BJPY2023A010), National Postdoctoral Researcher Assistance Program (GZC20232718) and Youth Project of the President’s Fund of Hefei Institutes of Physical Science(E36CGG2F).

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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

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

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

    Jie Zhang, Feng Xu, Xipeng Zhao, Zhicheng Yao, Jie Zhang & Shenglan Zhou

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

    Bin Chen

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

    Lijun Wu

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Contributions

J.Zhang: Conceptualization, data curation, formal analysis, validation, investigation, visualization, writing-original draft, writing-review and editing. B.Chen, F.Xu, R.Wang, X.Zhao, Z.Yao, J.Zhang, S.Zhou: investigation. A.Xu, L.Wu: Project administration, supervision. G.Zhao: Conceptualization, resources, data curation, formal analysis, supervision, funding acquisition, investigation, project administration, writing-review, and editing.

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Correspondence to Guoping Zhao.

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Zhang, J., Chen, B., Xu, F. et al. Phospho-TRIM21 orchestrates RPA2 ubiquitination switch to promote homologous recombination and tumor radio/chemo-resistance. Oncogene 44, 1106–1117 (2025). https://doi.org/10.1038/s41388-025-03288-1

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