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Transcriptional control and signal transduction, cell cycle

MAPK/ERK2 phosphorylates ERG at serine 283 in leukemic cells and promotes stem cell signatures and cell proliferation

Leukemia volume 30, pages 1552–1561 (2016)Cite this article

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Abstract

Aberrant ERG (v-ets avian erythroblastosis virus E26 oncogene homolog) expression drives leukemic transformation in mice and high expression is associated with poor patient outcomes in acute myeloid leukemia (AML) and T-acute lymphoblastic leukemia (T-ALL). Protein phosphorylation regulates the activity of many ETS factors but little is known about ERG in leukemic cells. To characterize ERG phosphorylation in leukemic cells, we applied liquid chromatography coupled tandem mass spectrometry and identified five phosphorylated serines on endogenous ERG in T-ALL and AML cells. S283 was distinct as it was abundantly phosphorylated in leukemic cells but not in healthy hematopoietic stem and progenitor cells (HSPCs). Overexpression of a phosphoactive mutant (S283D) increased expansion and clonogenicity of primary HSPCs over and above wild-type ERG. Using a custom antibody, we screened a panel of primary leukemic xenografts and showed that ERG S283 phosphorylation was mediated by mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling and in turn regulated expression of components of this pathway. S283 phosphorylation facilitates ERG enrichment and transactivation at the ERG +85 HSPC enhancer that is active in AML and T-ALL with poor prognosis. Taken together, we have identified a specific post-translational modification in leukemic cells that promotes progenitor proliferation and is a potential target to modulate ERG-driven transcriptional programs in leukemia.

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Acknowledgements

We thank the staff at the Mark Wainwright analytical centre for mass spectrometry and for cell sorting analyses and the donors and staff of the cord blood bank at the Prince of Wales hospital for CBs. This work was funded by the National Health and Medical Research Council (Australia), Anthony Rothe Memorial Trust (to JAIT and DB) and the Cancer Institute of NSW. YH received scholarships from UNSW Australia and the Translational Cancer Research Network (UNSW, Australia). JWHW is a Future Fellow of the Australian Research Council.

Author contributions

YH, JAIT, MLT, KK, DB, VC, ENG and AB performed experiments and analyzed data. JO, SRT, KLM, JWHW and JEP analyzed data. SS, RBL, KLM and JHB provided essential reagents. YH, JAIT, JWHW and JEP wrote the paper.

Author information

Author notes

  1. M L Tursky

    Present address: 8Current address: Blood, Stem Cells and Cancer Research, St Vincent’s Centre for Applied Medical Research, St Vincent’s Hospital, Sydney, New South Wales, Australia.,

  2. Y Huang and J A I Thoms: These authors contributed equally to this work.

Authors and Affiliations

  1. Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia

    Y Huang, J A I Thoms, M L Tursky, K Knezevic, D Beck, V Chandrakanthan, J W H Wong & J E Pimanda

  2. Children’s Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia

    M L Tursky, S Suryani, R B Lock & K L MacKenzie

  3. School of Mathematics and Statistics, UNSW Australia, Sydney, New South Wales, Australia

    J Olivier

  4. Department of Chemistry, University of Virginia, Charlottesville, VA, USA

    A Boulton & J H Bushweller

  5. School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia

    E N Glaros & S R Thomas

  6. Department of Hematology, Prince of Wales Hospital, Sydney, New South Wales, Australia

    J E Pimanda

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  1. Y Huang
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Huang, Y., Thoms, J., Tursky, M. et al. MAPK/ERK2 phosphorylates ERG at serine 283 in leukemic cells and promotes stem cell signatures and cell proliferation. Leukemia 30, 1552–1561 (2016). https://doi.org/10.1038/leu.2016.55

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