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Poly(ADP-ribosyl)ation of OVOL2 regulates aneuploidy and cell death in cancer cells

Oncogene volume 38, pages 2750–2766 (2019)Cite this article

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Abstract

Poly(ADP-ribosyl)ation (PARylation) is a post-translational modification by which poly ADP-ribose (PAR) polymers are covalently added to proteins through a PAR polymerase (PARP). Here, using proteomic approach, we identify the transcriptional regulator, OVOL2, is a novel substrate of PARP1 and can be PARylated at residues Lysine 145, Lysine 176, and Lysine 212 within its C2H2 zinc finger domains. Overexpression of PARylated OVOL2 alters cell morphology and induces lagging chromosomes and aneuploidy. To define the underlying molecular mechanism by which OVOL2 induces abnormal cell cycle and centrosome amplification, we uncover that the OVOL2 elevates the protein levels of Cyclin E by enhancing its stability. Furthermore, we identify Skp2, the E3 ubiquitin ligase of Cyclin E, as a direct target of PARylated OVOL2. Using ChIP assay, the OVOL2 binding site on the promoter region of Skp2 is mapped. To further explore the physiological effect, we show that PARylated OVOL2 can induce cell death. Furthermore, to investigate PARylated OVOL2 function in vivo, we further develop a null-mice xenograft model and generate MMTV-PyVT transgenic mice and monitor the effect of wild-type OVOL2 and non-PARylated OVOL2-3K/A mutants on tumor progression. Consistently, overexpression of wild-type OVOL2 in both null-mice xenograft and MMTV-PyVT transgenic mice displays significantly reduction of tumor progression, respectively, further indicating that the function of OVOL2 as a tumor suppressor in vivo is highly regulated by PARylation. Taken together, our study sheds new light on PARP1-induced PARylation as a critical event in the OVOL2-mediated regulation of chromosomal integrity and suppression of cancer cells growth.

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Acknowledgements

We thank all members in the Y-C Liou laboratory for their support and valuable discussion. We would also like to thank the Center of BioImaging Sciences (CBIS) for their technical support on confocal microscopy, and Flow Cytometry Laboratory in Yong Loo Lin School of Medicine at the National University of Singapore for FACS assay. In addition, the authors thank Protein Proteomic Center (PPC) for the technical support on protein identification. We particularly thank Dr. Boon Chuan Low for kindly providing several reagents and helpful discussions and suggestions. In addition, we thank Dr. Kyunghee Lee for helping imaging and cell cycle assay. This work was supported in part by MOE grants Tier 2, R-154-000-A61-112 and Tier 1, A15-114 from the Ministry of Education, Singapore to Y.-C.L; grants from the National Natural Science Foundation of China (Grant Numbers U1705284, 81772958) to B.-A.L.

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  1. These authors contributed equally: Rui Zhang, Jing-Jing Hong

Authors and Affiliations

  1. Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, 117543, Singapore

    Rui Zhang, Qiaoyun Yang, Chin-Tong Ong & Yih-Cherng Liou

  2. State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, 361005, Xiamen, Fujian, People’s Republic of China

    Jing-Jing Hong & Bo-An Li

  3. Temasek Life Sciences Laboratory, National University of Singapore, Singapore, 117604, Singapore

    Chin-Tong Ong

  4. NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore, 117573, Singapore

    Yih-Cherng Liou

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Zhang, R., Hong, JJ., Yang, Q. et al. Poly(ADP-ribosyl)ation of OVOL2 regulates aneuploidy and cell death in cancer cells. Oncogene 38, 2750–2766 (2019). https://doi.org/10.1038/s41388-018-0615-3

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