title: Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes.
abstract: STAT5 is constitutively phosphorylated in leukemic cells in approximately 70% of acute myeloid leukemia (AML) patients. To identify kinase candidates potentially responsible for STAT5 phosphorylation, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) mass spectrometry to detect phosphoproteins in AML cell lines. We established TEL-ARG and BCR-ABL fusion proteins as the mechanism underlying STAT5 phosphorylation in HT-93 and KBM-3 cells, respectively. In addition, we identified a JAK2 pseudokinase domain mutation in HEL cells and using siRNA downregulation, established JAK2 as the kinase responsible for phosphorylating STAT5. This study illustrates the benefit of LC-MS/MS mass spectrometry and siRNA for the identification of novel targets and mutations.
authors.affiliation: Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, USA. waltersd@ohsu.edu
authors.forename: Denise K
authors.initials: DK
authors.lastname: Walters
authors.forename: Valerie L
authors.initials: VL
authors.lastname: Goss
authors.forename: Eric P
authors.initials: EP
authors.lastname: Stoffregen
authors.forename: Ting-Lei
authors.initials: TL
authors.lastname: Gu
authors.forename: Kimberly
authors.initials: K
authors.lastname: Lee
authors.forename: Julie
authors.initials: J
authors.lastname: Nardone
authors.forename: Laura
authors.initials: L
authors.lastname: McGreevey
authors.forename: Michael C
authors.initials: MC
authors.lastname: Heinrich
authors.forename: Michael W
authors.initials: MW
authors.lastname: Deininger
authors.forename: Roberto
authors.initials: R
authors.lastname: Polakiewicz
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Druker
mesh_terms: D002273:Carcinogens / Q000378:metabolism; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000235:genetics; D040901:Proteomics
chemical_list: D002273:Carcinogens; D009363:Neoplasm Proteins