title: Necessity of tyrosine 719 and phosphatidylinositol 3'-kinase-mediated signal pathway in constitutive activation and oncogenic potential of c-kit receptor tyrosine kinase with the Asp814Val mutation.
abstract: Substitution of valine (Val) for aspartic acid (Asp) at codon 814 constitutively activates murine c-kit receptor tyrosine kinase (KIT), and Asp816Val mutation, corresponding to murine Asp814Val mutation, is found in patients with mastocytosis and acute myelocytic leukemia. However, the signal transduction pathways responsible for oncogenesis by the Asp814Val mutant (KIT(Val814)) are not fully understood. To examine the oncogenic signal transduction of KIT(Val814), we converted 20 tyrosine (Tyr) residues to phenylalanine (Phe) in the cytoplasmic domain of KIT(Val814) or deleted the C-terminal region containing 2 other tyrosine residues (Del). Among various KIT(Val814)- derived mutants, KIT(Val814-Tyr719Phe) and KIT(Val814-Del) severely impaired receptor tyrosine phosphorylation and association with the p85 subunit of phosphatidylinositol 3'-kinase (p85 (PI3-K)). Moreover, KIT(Val814-Tyr719Phe) and KIT(Val814-Del) failed to induce ligand-independent growth in Ba/F3 cells, indicating that Tyr719, the binding site for p85(PI3-K), and the C-terminal region are indispensable for factor-independent growth by KIT(Val814). Although the C-terminal region was also required for ligand-dependent growth by wild-type KIT (KIT(WT)), the Tyr719Phe substitution had negligible effects on ligand-dependent growth by KIT(WT). Furthermore, dominant-negative PI3-K significantly inhibited ligand-independent growth by KIT(Val814). These results demonstrate that Tyr719 is crucial for constitutive activation of KIT(Val814), but not for the ligand-induced activation of KIT(WT), and that the downstream signaling of PI3-K plays an important role in ligand-independent growth and tumorigenicity by KIT(Val814), thereby suggesting that KIT(Val814) is a unique activating mutation that leads to a distinguishable function from the effects of KIT(WT).
authors.affiliation: Department of Pathology, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Itaru
authors.initials: I
authors.lastname: Matsumura
authors.forename: Tohru
authors.initials: T
authors.lastname: Tsujimura
authors.forename: Dae-Ki
authors.initials: DK
authors.lastname: Kim
authors.forename: Hideki
authors.initials: H
authors.lastname: Ogihara
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Ikeda
authors.forename: Shuji
authors.initials: S
authors.lastname: Ueda
authors.forename: Masao
authors.initials: M
authors.lastname: Mizuki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sugahara
authors.forename: Hirohiko
authors.initials: H
authors.lastname: Shibayama
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Kitamura
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Kanakura
mesh_terms: D000818:Animals; D002455:Cell Division; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D004789:Enzyme Activation; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship; D014162:Transfection; D014443:Tyrosine / Q000235:genetics
chemical_list: D014443:Tyrosine; D019869:Phosphatidylinositol 3-Kinases; D019009:Proto-Oncogene Proteins c-kit