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The protection of UCK2 protein stability by GART maintains pyrimidine salvage synthesis for HCC growth under glucose limitation

Oncogene volume 44, pages 1078–1092 (2025)Cite this article

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Abstract

Overexpression of uridine-cytidine kinase 2 (UCK2), a key enzyme in the pyrimidine salvage pathway, is implicated in human cancer development, while its regulation under nutrient stress remains to be investigated. Here, we show that under glucose limitation, AMPK phosphorylates glycinamide ribonucleotide formyltransferase (GART) at Ser440, and this modification facilitates its interaction with UCK2. Through its binding to UCK2, GART generates tetrahydrofolate (THF) and thus inhibits the activity of integrin-linked kinase associated phosphatase (ILKAP) for removing AKT1-mediated UCK2-Ser254 phosphorylation under glucose limitation, in which dephosphorylation of UCK2-Ser254 tends to cause Trim21-mediated UCK2 polyubiquitination and degradation. In this way, both UCK2 binding ability and THF producing catalytic activity of GART protect protein stability of UCK2 and pyrimidine salvage synthesis, and sustain tumor cell growth under glucose limitation. In addition, UCK2-Ser254 phosphorylation level displays a positive relationship with GART-Ser440 phosphorylation level and its enhancement is correlated with poor prognosis of human hepatocellular carcinoma (HCC) patients. These findings reveal a non-canonical role of GART in regulating pyrimidine salvage synthesis under nutrient stress, and raise the potential for alternative treatments in targeting pyrimidine salvage-dependent tumor growth.

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Fig. 1: GART sustains pyrimidine salvage synthesis under glucose limitation.
Fig. 2: GART maintains UCK2 protein stability under glucose limitation.
Fig. 3: GART positively regulates AKT1-mediated UCK2 phosphorylation and UCK2 protein stability.
Fig. 4: AMPK phosphorylates GART and promotes GART-UCK2 interaction.
Fig. 5: The interaction of GART with UCK2 is required for maintaining UCK2 protein stability.
Fig. 6: The enzymatic activity of GART inhibits ILKAP phosphatase activity against UCK2 phosphorylation.
Fig. 7: The positive regulation of UCK2 by GART-Ser440 phosphorylation facilitates liver cancer development.

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Data availability

Supplemental Figures and Table are available in the Supplemental Figures and Table file. The mass spectrometry proteomics data can be accessed at the iProX site.

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Acknowledgements

We thank the staff members of the Large-scale Protein Preparation System at the National Facility for Protein Science in Shanghai (NFPS), Shanghai Advanced Research Institute, Chinese Academy of Sciences, China for providing technical support and assistance in data collection and analysis.

Funding

This work was supported by National Key R&D Program of China (2023YFA1802004 to Y. Jiang), National Nature Science Foundation of China (82273230 to Q. Zhao; 81972586 to Y. Jiang).

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Author notes

  1. These authors contributed equally: Nannan Sha, Bei Zhou.

Authors and Affiliations

  1. Department of Liver Surgery and Shanghai Cancer Institute, State Key Laboratory of Systems Medicine for Cancer, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Nannan Sha, Bei Zhou, Guofang Hou, Zhifeng Xi, Wang Wang, Man Yan, Jing He, Yue Zhou, Qiang Xia & Qin Zhao

  2. Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism, School of Pharmacy, East China University of Science and Technology, Shanghai, China

    Yuhui Jiang

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Contributions

This study was conceived by Q. Zhao, Y. Jiang and Q. Xia; Q. Zhao and Y. Jiang designed the study; Q. Zhao, N. Sha, B. Zhou, G. Hou, Z. Xi, W. Wang, M. Yan, J. He, Y. Zhou performed experiments; Q. Zhao and Y. Jiang wrote the paper with comments from all authors.

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Correspondence to Qiang Xia, Yuhui Jiang or Qin Zhao.

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The authors confirm that all methods were performed in accordance with the relevant guidelines and regulations. This study was reviewed and approved by Shanghai Jiaotong University School of Medicine, Renji Hospital Ethics Committee (KY2024-152-B) with written informed consents from all the patients.

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Sha, N., Zhou, B., Hou, G. et al. The protection of UCK2 protein stability by GART maintains pyrimidine salvage synthesis for HCC growth under glucose limitation. Oncogene 44, 1078–1092 (2025). https://doi.org/10.1038/s41388-025-03274-7

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