Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes.
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.
Walters
Denise K
DK
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
Goss
Valerie L
VL
Stoffregen
Eric P
EP
Gu
Ting-Lei
TL
Lee
Kimberly
K
Nardone
Julie
J
McGreevey
Laura
L
Heinrich
Michael C
MC
Deininger
Michael W
MW
Polakiewicz
Roberto
R
Druker
Brian J
BJ
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
D002273:Carcinogens; D009363:Neoplasm Proteins