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Phosphorylation of CBX8 by PKD1 suppresses PRC1 activity and promotes cell senescence

Oncogene volume 45, pages 398–413 (2026)Cite this article

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Abstract

The Polycomb group (PcG) protein chromobox 8 (CBX8) is the subunit of Polycomb repressive complex 1 (PRC1) and recognizes the trimethylation of histone H3 on Lysine 27 (H3K27me3), and coordinates with PRC2 complex to function as an epigenetic gene silencer. CBX8 plays a key role in cell proliferation, stem cell biology, cell senescence, and cancer development. However, the post-translational modifications of CBX8 remain poorly understood. Here, we report that protein kinase D1 (PKD1) interacts and phosphorylates CBX8 at Thr234 and Ser256 /311 residues. PKD1-mediated CBX8 phosphorylation at Thr234 reduced its expression level by promoting its ubiquitination-mediated degradation, whereas Ser256/311 phosphorylation decreased CBX8 binding to other PRC1 components BMI1 and RING1A. Overall, CBX8 phosphorylation by PKD1 impaired PRC1 complex integrity and activity, mitigated H2AK119ub1 level, caused the upregulation of multiple target genes repressed by CBX8, and decreased CBX8, H2AK119ub1, and H3K27me3 enrichment at INK4A/ARF locus, thereby derepressing p16INK4A and facilitating cellular senescence. Collectively, these results suggest that PKD1-mediated CBX8 phosphorylation at T234 and S256/311 is a key mechanism governing CBX8 function, including cell senescence.

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Fig. 1: PKD1 associates with CBX8 and phosphorylates CBX8.
Fig. 2: PKD1 phosphorylates CBX8 at Thr234, Ser256 and Ser311 sites.
Fig. 3: CBX8-T234 phosphorylation by PKD1 promotes its degradation via the ubiquitin-proteasome pathway.
Fig. 4: PKD1-mediated CBX8 phosphorylation disrupts PRC1 complex integrity.
Fig. 5: CBX8 phosphorylation by PKD1 suppresses H2AK119ub1 level.
Fig. 6: CBX8 phosphorylation by PKD1 abrogates CBX8-mediated silencing of PRC1 target genes.
Fig. 7: PKD1-mediated CBX8 phosphorylation reduces the binding enrichment of CBX8, H2AK119ub1, and H3K27me3 at the INK4A locus.
Fig. 8: PKD1-mediated CBX8 phosphorylation upregulates p16INK4A expression and promotes cell senescence.

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Data availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD045225. The website is: http://www.ebi.ac.uk/pride.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (31871382 and 32070733) and from the Ministry of Science and Technology of the People’s Republic of China (2018YFC2000102).

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Authors and Affiliations

  1. Peking University Research Center on Aging, Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Protein Posttranslational Modifications and Cell Function, Department of Integration of Chinese and Western Medicine, School of Basic Medical Science, Peking University, Beijing, 100191, China

    Zhiqiang Fang, Doudou Liu, Yuanyuan Su, Fengxin Hao, Ruodong Qin, Guodong Li & Jun Chen

  2. Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China

    Doudou Liu

  3. Xiamen Key Laboratory of Precision Medicine for Cardiovascular Disease, Xiamen, China

    Yuanyuan Su

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Contributions

JC conceived the study. JC, ZQF, YYS, and DDL designed the experiments. ZQF performed most of the experiments; YYS conducted Figs. 1A–C, S1, 2C, S2H and S7A, B, D; DDL performed Fig. 1D, F, SILAC experiment, and Fig. 2E, F; FXH performed RNA-seq experiment; RDQ conducted AlphaFold3 prediction. GDL provided resources. ZQF analyzed most of the data and prepared figures. JC, ZQF, and YYS wrote the manuscript.

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Correspondence to Jun Chen.

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Fang, Z., Liu, D., Su, Y. et al. Phosphorylation of CBX8 by PKD1 suppresses PRC1 activity and promotes cell senescence. Oncogene 45, 398–413 (2026). https://doi.org/10.1038/s41388-025-03657-w

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