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Constitutively activating mutations of c-kit receptor tyrosine kinase
confer factor-independent growth and tumorigenicity of factor-dependent
hematopoietic cell lines
H Kitayama, Y Kanakura, T Furitsu, T Tsujimura, K Oritani, H Ikeda, H Sugahara, H Mitsui, Y Kanayama and Y Kitamura
Second Department of Internal Medicine, Osaka University Medical School,
Japan.
The c-kit receptor tyrosine kinase (KIT) is activated upon ligand binding,
thereby leading to a variety of signaling events that play a fundamental
role in hematopoiesis. In addition to ligand-dependent activation, we have
previously shown that KIT is constitutively activated in a
ligand-independent manner by two point mutations, Val- 559-->Gly (G559)
mutation in the juxtamembrane domain and Asp-814-->Val (V814) mutation
in the phosphotransferase domain. To investigate the biochemical
consequence and biologic significance of these mutations, retroviral
vectors encoding KITG559 or KITV814 were introduced into murine pro-B-type
Ba/F3 cells and myeloid FDC-P1 cells, both of which require interleukin-3
(IL-3) for their growth and survival. In the cells, KITG559 or KITV814 were
found to be constitutively phophorylated on tyrosine in the absence of stem
cell factor (SCF) that is a ligand for KIT. Chemical cross-linking analysis
showed that a substantial fraction of the phosphorylated KITG559 underwent
dimerization even in the absence of SCF, whereas the phosphorylated KITV814
did not, suggesting the distinct mechanisms underlying constitutive
activation of KIT by G559 and V814 mutations. Furthermore, the cells
expressing either KITG559 or KITV814 were found to show a
factor-independent growth, whereas the cells expressing wild-type KIT
(KITWT) proliferated in response to SCF as well as IL-3. Moreover,
subcutaneous injection of Ba/F3 cells expressing KITG559 or KITV814 into
nude mice resulted in production of large tumors at all sites of the
injection within 2 weeks, and all nude mice quickly succumbed to leukemia
and died. These results suggest that, although the mechanisms underlying
constitutive activation of KITG559 or KITV814 may be different, both of the
activating mutations have a function to induce a factor-independent and
tumorigenic phenotype. Also, the data of this study raise the possibility
that the constitutively activating mutations of c-kit may play a causal
role in development of hematologic malignancies.
Volume 85,
Issue 3,
pp. 790-798,
02/01/1995
Copyright © 1995 by The American Society of Hematology
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