Activating mutations of the c-kit proto-oncogene in a human mast cell leukemia cell line

Yuzuru Kanakura, Takuma Furitsu, Tohru Tsujimura, Joseph H. Butterfield, Leonie K. Ashman, Hirokazu Ikeda, Hitoshi Kitayama, Yoshio Kanayama, Yuji Matsuzawa, Yukihiko Kitamura

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The c-kit proto-oncogene encodes a receptor tyrosine kinase that is known to play a crucial role in mast cell growth and differentiation. In a human mast cell leukemia cell line (HMC-1), KitR was found to be constitutively phosphorylated on tyrosine, activated and associated with phosphatidylinositol 3-kinase (PI3K) in the absence of autocrine production of SCF. Sequencing of c-kit cDNA revealed that c-kit genes of HMC-1 cells were composed of a normal, wild-type allele and a mutant allele with two point mutations in codon 560 and codon 816, resulting in intracellular amino acid substitutions of Gly-560 for Val and Val-816 for Asp, respectively. Murine c-kit mutants encoding Gly-559 and/or Val-814, corresponding to human Gly-560 and/or Val-816, were constructed by site-directed mutagenesis and expressed in cells of a human embryonic kidney cell line (293T). In the transfected cells, KitR (Gly-559 + Val-814) and KitR (Val-814) were strikingly phosphorylated on tyrosine and activated in the absence of SCF, whereas tyrosine phosphorylation and activation of KitR (Gly-559) or wild-type KitR was modest or little, respectively. These results suggest that constitutive activation of KitR in HMC-1 results from the activating mutations of c-kit gene, and raise the possibility that the activating mutations, particularly at codon 814 of murine c-kit or at codon 816 of human c-kit, may participate in oncogenesis of mast cells.

Original languageEnglish (US)
JournalLeukemia
Volume8
Issue numberSUPPL.1
StatePublished - 1994
Externally publishedYes

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Mast-Cell Leukemia
Proto-Oncogenes
Codon
Tyrosine
Cell Line
Mutation
Mast Cells
Alleles
Phosphatidylinositol 3-Kinase
Receptor Protein-Tyrosine Kinases
Amino Acid Substitution
Viperidae
Site-Directed Mutagenesis
Point Mutation
Genes
Cell Differentiation
Carcinogenesis
Complementary DNA
Phosphorylation
Kidney

ASJC Scopus subject areas

  • Cancer Research
  • Hematology

Cite this

Kanakura, Y., Furitsu, T., Tsujimura, T., Butterfield, J. H., Ashman, L. K., Ikeda, H., ... Kitamura, Y. (1994). Activating mutations of the c-kit proto-oncogene in a human mast cell leukemia cell line. Leukemia, 8(SUPPL.1).

Activating mutations of the c-kit proto-oncogene in a human mast cell leukemia cell line. / Kanakura, Yuzuru; Furitsu, Takuma; Tsujimura, Tohru; Butterfield, Joseph H.; Ashman, Leonie K.; Ikeda, Hirokazu; Kitayama, Hitoshi; Kanayama, Yoshio; Matsuzawa, Yuji; Kitamura, Yukihiko.

In: Leukemia, Vol. 8, No. SUPPL.1, 1994.

Research output: Contribution to journalArticle

Kanakura, Y, Furitsu, T, Tsujimura, T, Butterfield, JH, Ashman, LK, Ikeda, H, Kitayama, H, Kanayama, Y, Matsuzawa, Y & Kitamura, Y 1994, 'Activating mutations of the c-kit proto-oncogene in a human mast cell leukemia cell line', Leukemia, vol. 8, no. SUPPL.1.
Kanakura Y, Furitsu T, Tsujimura T, Butterfield JH, Ashman LK, Ikeda H et al. Activating mutations of the c-kit proto-oncogene in a human mast cell leukemia cell line. Leukemia. 1994;8(SUPPL.1).
Kanakura, Yuzuru ; Furitsu, Takuma ; Tsujimura, Tohru ; Butterfield, Joseph H. ; Ashman, Leonie K. ; Ikeda, Hirokazu ; Kitayama, Hitoshi ; Kanayama, Yoshio ; Matsuzawa, Yuji ; Kitamura, Yukihiko. / Activating mutations of the c-kit proto-oncogene in a human mast cell leukemia cell line. In: Leukemia. 1994 ; Vol. 8, No. SUPPL.1.
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AU - Ashman, Leonie K.

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