title: MAPK/ERK2 phosphorylates ERG at serine 283 in leukemic cells and promotes stem cell signatures and cell proliferation.
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.
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: J A I
authors.initials: JA
authors.lastname: Thoms
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.|Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: M L
authors.initials: ML
authors.lastname: Tursky
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: K
authors.initials: K
authors.lastname: Knezevic
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: D
authors.initials: D
authors.lastname: Beck
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: V
authors.initials: V
authors.lastname: Chandrakanthan
authors.affiliation: Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Suryani
authors.affiliation: School of Mathematics and Statistics, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: J
authors.identifier: 0000-0002-3144-4507
authors.initials: J
authors.lastname: Olivier
authors.affiliation: Department of Chemistry, University of Virginia, Charlottesville, VA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Boulton
authors.affiliation: School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: E N
authors.initials: EN
authors.lastname: Glaros
authors.affiliation: School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: S R
authors.initials: SR
authors.lastname: Thomas
authors.affiliation: Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: R B
authors.initials: RB
authors.lastname: Lock
authors.affiliation: Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: K L
authors.initials: KL
authors.lastname: MacKenzie
authors.affiliation: Department of Chemistry, University of Virginia, Charlottesville, VA, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Bushweller
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: J W H
authors.initials: JW
authors.lastname: Wong
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, UNSW Australia, Sydney, New South Wales, Australia.|Department of Hematology, Prince of Wales Hospital, Sydney, New South Wales, Australia.
authors.forename: J E
authors.initials: JE
authors.lastname: Pimanda
mesh_terms: D001665:Binding Sites; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006412:Hematopoietic Stem Cells; D006801:Humans; D007938:Leukemia / Q000473:pathology*; D015470:Leukemia, Myeloid, Acute / Q000473:pathology; D020935:MAP Kinase Signaling System / Q000502:physiology*; D010766:Phosphorylation; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology; D011499:Protein Processing, Post-Translational; D012694:Serine / Q000378:metabolism; D000071230:Transcriptional Regulator ERG / Q000378:metabolism; D059467:Transcriptome
chemical_list: C072312:ERG protein, human; D000071230:Transcriptional Regulator ERG; D012694:Serine