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Cell-cycle-dependent phosphorylation of RRM1 ensures efficient DNA replication and regulates cancer vulnerability to ATR inhibition

Oncogene volume 39, pages 5721–5733 (2020)Cite this article

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Abstract

Ribonucleotide reductase (RNR) catalyzes the rate-limiting step of de novo synthesis of deoxyribonucleotide triphosphates (dNTPs) building blocks for DNA synthesis, and is a well-recognized target for cancer therapy. RNR is a heterotetramer consisting of two large RRM1 subunits and two small RRM2 subunits. RNR activity is greatly stimulated by transcriptional activation of RRM2 during S/G2 phase to ensure adequate dNTP supply for DNA replication. However, little is known about the cell-cycle-dependent regulation of RNR activity through RRM1. Here, we report that RRM1 is phosphorylated at Ser 559 by CDK2/cyclin A during S/G2 phase. And this S559 phosphorylation of RRM1enhances RNR enzymatic activity and is required for maintaining sufficient dNTPs during normal DNA replication. Defective RRM1 S559 phosphorylation causes DNA replication stress, double-strand break, and genomic instability. Moreover, combined targeting of RRM1 S559 phosphorylation and ATR triggers lethal replication stress and profound antitumor effects. Thus, this posttranslational phosphorylation of RRM1 provides an alternative mechanism to finely regulating RNR and therapeutic opportunities for cancer treatment.

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Fig. 1: RRM1 is phosphorylated at ser/thr residues during S and early G2 phase of the cell-cycle.
Fig. 2: RRM1 is phosphorylated at serine 559 by CDK2/cyclin A.
Fig. 3: S559 phosphorylation of RRM1 stimulates RNR enzymatic activity.
Fig. 4: Defective RRM1 phosphorylation on S559 induces DNA replication stress.
Fig. 5: S559 phosphorylation of RRM1 is required to maintain genome stability.
Fig. 6: Inhibition of CDK2-RRM1 phosphorylation sensitizes human lung cancer cells to ATR inhibitor VE822.
Fig. 7: A schematic illustration of the role of RRM1 phosphorylation in DNA replication stress.

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Acknowledgements

We are grateful to Stephanie Jamison for the English editing.

Funding

This work was supported by Natural Science Foundation of Guangdong Province of China (2019A1515011247 to GC), Guangzhou Municipal Science and Technology Program of China (202002030451 to GC), Fundamental Research Funds for the Central Universities (21620421 to GC) and National Natural Science Foundation of China (81472793 and 81672895 to DW).

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  1. These authors contributed equally: Zhen Shu, Zhen Li, Huanhuan Huang

Authors and Affiliations

  1. Department of Medical Biochemistry and Molecular Biology, School of Medicine, Jinan University, Guangzhou, 510632, China

    Zhen Shu, Zhen Li, Yan Chen, Jun Fan, Li Yu, Zhihui Wu, Ling Tian & Guo Chen

  2. Department of Radiation Oncology, Emory University School of Medicine, Atlanta, GA, 30322, USA

    Zhen Shu, Changyong Wei & Zhongqiu Xie

  3. Biomedical Translational Research Institute, Jinan University, Guangzhou, 510632, China

    Zhen Li, Qi Feng, Hao Sheng, Guangqiang Li & Changliang Shan

  4. Department of Oncology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China

    Huanhuan Huang & Dongping Wei

  5. Department of Pathophysiology and Pharmacology, School of Medicine, Jinan University, Guangzhou, 510632, China

    Qi Qi & Shuang Peng

  6. Key Laboratory of Immune Microenvironment and Disease of the Ministry of Education, Tianjin Medical University, Tianjin, 300070, China

    Xiaobo Li

  7. State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China

    Changliang Shan

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Shu, Z., Li, Z., Huang, H. et al. Cell-cycle-dependent phosphorylation of RRM1 ensures efficient DNA replication and regulates cancer vulnerability to ATR inhibition. Oncogene 39, 5721–5733 (2020). https://doi.org/10.1038/s41388-020-01403-y

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