Abstract
Tyrosine kinases are aberrantly activated in numerous malignancies, including acute myeloid leukemia (AML). To identify tyrosine kinases activated in AML, we developed a screening strategy that rapidly identifies tyrosine-phosphorylated proteins using mass spectrometry. This allowed the identification of an activating mutation (A572V) in the JAK3 pseudokinase domain in the acute megakaryoblastic leukemia (AMKL) cell line CMK. Subsequent analysis identified two additional JAK3 alleles, V722I and P132T, in AMKL patients. JAK3(A572V), JAK3(V722I), and JAK3(P132T) each transform Ba/F3 cells to factor-independent growth, and JAK3(A572V) confers features of megakaryoblastic leukemia in a murine model. These findings illustrate the biological importance of gain-of-function JAK3 mutations in leukemogenesis and demonstrate the utility of proteomic approaches to identifying clinically relevant mutations.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Alleles
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Animals
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Apoptosis / drug effects
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Benzamides
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Cell Line, Tumor
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Cell Proliferation / drug effects
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Cell Survival / drug effects
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Humans
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Imatinib Mesylate
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Janus Kinase 2
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Janus Kinase 3
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K562 Cells
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Leukemia, Experimental / genetics*
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Leukemia, Experimental / metabolism
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Leukemia, Experimental / pathology
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Leukemia, Megakaryoblastic, Acute / genetics*
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Leukemia, Megakaryoblastic, Acute / metabolism
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Leukemia, Megakaryoblastic, Acute / pathology
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Mice
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Mice, Inbred C57BL
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Models, Molecular
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Mutant Proteins / chemistry
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Mutant Proteins / genetics
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Mutant Proteins / metabolism
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Phosphorylation / drug effects
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Piperazines / pharmacology
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Protein Kinase Inhibitors / pharmacology
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Protein Structure, Tertiary / genetics
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Protein-Tyrosine Kinases / antagonists & inhibitors
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Protein-Tyrosine Kinases / genetics*
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Protein-Tyrosine Kinases / metabolism
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism
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Pyrimidines / pharmacology
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RNA, Small Interfering / genetics
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TYK2 Kinase
Substances
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Benzamides
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Mutant Proteins
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Piperazines
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Protein Kinase Inhibitors
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Proto-Oncogene Proteins
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Pyrimidines
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RNA, Small Interfering
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Imatinib Mesylate
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Protein-Tyrosine Kinases
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JAK2 protein, human
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JAK3 protein, human
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Jak2 protein, mouse
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Jak3 protein, mouse
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Janus Kinase 2
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Janus Kinase 3
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TYK2 Kinase
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TYK2 protein, human