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Protein kinase A-mediated phosphorylation regulates STAT3 activation and oncogenic EZH2 activity

Oncogene volume 37, pages 3589–3600 (2018)Cite this article

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Abstract

Polycomb repressive complex 2 (PRC2) member enhancer of zeste homolog 2 (EZH2) trimethylates histone H3 lysine 27 (H3K27me3), alters chromatin structure and contributes to epigenetic regulation of gene expression in normal and disease processes. Phosphorylation of EZH2 augmented EZH2 oncogenic activity in cancer but observations have been limited to threonine 350 (T350) and serine 21 (S21) residues by cyclin-dependent kinase 1 and protein kinase B, respectively. In addition, phosphorylation of the evolutionarily conserved T372 motif of EZH2 by p38 resulted in EZH2 interaction with Ying Yang 1 and promoted muscle stem cell differentiation. In the present study, we used epithelial ovarian cancer (OC) cells as a model to demonstrate that phosphorylation of EZH2 at T372 by protein kinase A (PKA) induced a dominant-negative EZH2 phenotype, inhibited OC cell proliferation and migration in vitro and decreased ovarian xenograft tumor growth in vivo. Phosphorylation of T372 by PKA enhanced the interaction between EZH2 and signal transducer and activator of transcription 3 (STAT3), and STAT3 binding to pT372-EZH2 reduced cellular levels of pSTAT3 and downregulated interleukin 6 receptor expression in OC. Furthermore, PKA-mediated pT372-EZH2 decreased ATP levels and altered mitochondrial gene expression, resulting in mitochondrial dysfunction and reduced OC cell growth. These findings demonstrate that PKA-mediated T372 phosphorylation reduces oncogenic EZH2 activity and reveal a novel role for pT372 in regulating EZH2 in OC and possibly other cancers.

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Acknowledgements

We thank Dr. Fang Fang and Jay Pilrose for assistance with animal studies, Dr. Scott Michaels, Dr. Doug Rusch and Aaron Buechlein (Center for Genomics and Bioinformatics, Indiana University) for RNA-seq data analysis, Dr. Haojie Huang for EZH2-pGEX-4T-1 vectors (University of Minnesota) and Dr. Jian Jian (UC Davis) for PGL2-3 × -SIEGAS STAT3 reporter plasmid. We thank Dr. Heather O’Hagan and Dr. Peter Hollenorst (Medical Sciences, Indiana University School of Medicine) for critically reading the manuscript and offering valuable suggestions. This work was made possible by funding from the National Cancer Institute (Awards CA13001 and CA182832), The V Foundation for Cancer Research Translational Grant, Walther Cancer Foundation (Indianapolis, IN) and the Doane and Eunice Dahl Wright Fellowship (Medical Sciences Program, Indiana University).

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Authors and Affiliations

  1. Molecular and Cellular Biochemistry Department, Indiana University, Bloomington, IN, 47405, USA

    Ali R. Özeş, Nick Pulliam & Kenneth P. Nephew

  2. Indiana University School of Medicine, Medical Sciences, Bloomington, IN, 47405, USA

    Ali R. Özeş, Nick Pulliam, Jessica Tang & Kenneth P. Nephew

  3. Department of Medical Biology and Genetics, Akdeniz University, Antalya, Turkey

    Mustafa G. Ertosun, Özlem Yılmaz, Ece Çopuroğlu & Osman N. Özeş

  4. Department of Obstetrics and Gynecology, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA

    Daniela Matei

  5. Departments of Cellular and Integrative Physiology and Obstetrics and Gynecology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA

    Kenneth P. Nephew

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  1. Ali R. Özeş
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Correspondence to Osman N. Özeş or Kenneth P. Nephew.

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Özeş, A.R., Pulliam, N., Ertosun, M.G. et al. Protein kinase A-mediated phosphorylation regulates STAT3 activation and oncogenic EZH2 activity. Oncogene 37, 3589–3600 (2018). https://doi.org/10.1038/s41388-018-0218-z

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