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TRIM28 interacts with EZH2 and SWI/SNF to activate genes that promote mammosphere formation

Oncogene volume 36, pages 2991–3001 (2017)Cite this article

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Abstract

Histone methyl transferase EZH2 (Enhancer of Zeste Homolog 2) is generally associated with H3K27 methylation and gene silencing, as a member of the polycomb repressor 2 (PRC2) complex. Immunoprecipitation and mass spectrometry of the EZH2–protein interactome in estrogen receptor positive, breast cancer-derived MCF7 cells revealed EZH2 interactions with subunits of chromatin remodeler SWI/SNF complex and TRIM28, which formed a complex with EZH2 distinct from PRC2. Unexpectedly, transcriptome profiling showed that EZH2 primarily activates, rather than represses, transcription in MCF7 cells and with TRIM28 co-regulates a set of genes associated with stem cell maintenance and poor survival of breast cancer patients. TRIM28 depletion repressed EZH2 recruitment to chromatin and expression of this gene set, in parallel with decreased CD44hi/CD24lo mammosphere formation. Mammosphere formation, inhibited by EZH2 depletion, was rescued by ectopic expression of EZH2 but not by TRIM28 expression or by EZH2 mutated at the region (pre-SET domain) of TRIM28 interaction. These results support PRC2-independent functions of EZH2 and TRIM28 in activation of gene expression that promotes mammary stem cell enrichment and maintenance.

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Acknowledgements

This work was supported in part by CPRIT RP110471 to MCB, WL and SYD. JL received support by a Gigli Family Endowed Scholarship of the UT-Graduate School of Biomedical Sciences at Houston, TX. AKJ was supported by The Laura and John Arnold Foundation and Odyssey Program fellowship. We thank Dr M-C Hung for the Myc-EZH2 plasmid, Dr J-I Park for Flag-EZH2 plasmids and Dr PR Potts for HA-TRIM28, GST-TRIM28 full-length and RBCC only plasmids. This study used the Science Park NGS Core, supported by CPRIT Core Facility Support Grant RP120348, and the Flow Cytometry and Cellular Imaging Facility, which is supported in part by the National Institutes of Health through MD Anderson's Cancer Center Support Grant CA016672.

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Authors and Affiliations

  1. Genes and Development Graduate Program, The University of Texas Graduate School of Biomedical Sciences at Houston, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    J Li & M C Barton

  2. Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    J Li, W Li, R L McCarthy, S A Stratton, S Y Dent, A K Jain & M C Barton

  3. Center for Cancer Epigenetics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    J Li, W Li, R L McCarthy, S A Stratton, S Y Dent, A K Jain & M C Barton

  4. Department of Surgery, University of Michigan School of Medicine, Ann Arbor, Michigan, USA

    Y Xi & W Zou

  5. Department of Molecular and Cellular Biology, Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, TX, USA

    W Li

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Li, J., Xi, Y., Li, W. et al. TRIM28 interacts with EZH2 and SWI/SNF to activate genes that promote mammosphere formation. Oncogene 36, 2991–3001 (2017). https://doi.org/10.1038/onc.2016.453

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