Abstract
The DNA damage response (DDR) is critical for maintaining cellular homeostasis and genome integrity. Mounting evidence has shown that posttranslational protein modifications play vital roles in the DDR. In this study, we showed that deubiquitinase OTUD6A is involved in the DDR and is important for maintaining genomic stability. Mechanistically, in response to DNA damage, the abundance of OTUD6A was increased; meanwhile, PP2A interacted with OTUD6A and dephosphorylated OTUD6A at sites S70/71/74, which promoted nuclear localization of OTUD6A. Subsequently, OTUD6A was recruited to the damage site, where it interacted with TopBP1 and blocked the interaction between TopBP1 and its ubiquitin E3 ligase UBR5, decreasing K48-linked polyubiquitination and increasing the stability of TopBP1. OTUD6A depletion impaired CHK1 S345 phosphorylation and blocked cell cycle progression under DNA replication stress. Consistently, knockout of OTUD6A rendered mice hypersensitive to irradiation, shortened survival, and inhibited tumor growth by regulating TopBP1 in xenografted nude mice. Moreover, OTUD6A is expressed at high levels in breast cancer, and OTUD6A overexpression promotes cell proliferation, migration and invasion, indicating that dysregulation of OTUD6A expression contributes to genomic instability and is associated with tumor development. In summary, this study demonstrates that OTUD6A plays a critical role in promoting tumor cell resistance to chemoradiotherapy by deubiquitinating and stabilizing TopBP1.
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Data availability
All data generated or analyzed during this study are included in this article and its supplementary data files and all original data are available from the corresponding authors upon request.
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Acknowledgements
We sincerely thank Prof. Lingqiang Zhang for providing OTUD6A cDNA, Prof. Jiadong Wang for providing the Myc-TopBP1 plasmid, Prof. Shimin Zhao for providing Myc-UBR5 plasmids, and Prof. Daochun Kong for providing materials used in EMSA experiment. We thank the National Center for Protein Sciences at Peking University, particularly Liying Du, Huan Yang, Dong Liu, Qi Zhang, Hongxia Lv, LiQin Fu, and Guilan Li, for technical help. We also appreciate the assistance of Xiaochen Li and Siying Qin from the Core Facilities of Life Sciences at Peking University for their assistance with microscopic imaging. This work was supported by the National Natural Science Foundation of China (82130081 and 81730080) and the Natural Science Foundation of Beijing Municipality (5212008).
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YZ designed and performed the experiments, analyzed the data, and wrote the manuscript. XH, DZ, and JL performed the experiments. MW analyzed the pathway data and performed experiments. SY provided technical support for metaphase spread assays. YT performed the cancer cell injection into mice. XZ designed and supervised this study and wrote the manuscript.
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Zhao, Y., Huang, X., Zhu, D. et al. Deubiquitinase OTUD6A promotes breast cancer progression by increasing TopBP1 stability and rendering tumor cells resistant to DNA-damaging therapy. Cell Death Differ 29, 2531–2544 (2022). https://doi.org/10.1038/s41418-022-01036-6
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DOI: https://doi.org/10.1038/s41418-022-01036-6
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