Phosphatidylinositol 3 kinase contributes to the transformation of hematopoietic cells by the D816V c-Kit mutant

Ru Ju Chian, Sonia Young, Alla Danilkovitch-Miagkova, Lars Rönnstrand, Edward Leonard, Petranel Ferrao, Leonie Ashman, Diana Linnekin

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

Stem cell factor (SCF) binds the receptor tyrosine kinase c-Kit and is critical for normal hematopoiesis. Substitution of valine for aspartic acid 816 (D816V) constitutively actives human c-Kit, and this mutation is found in patients with mastocytosis, leukemia, and germ cell tumors. Immortalized murine progenitor cells (MIHCs) transduced with wild-type c-Kit proliferate in response to SCF, whereas cells expressing D816V c-Kit (MIHC-D816V) are factor-independent and tumorigenic. However, the mechanisms mediating transformation by D816V c-Kit are unknown. The objective of this study was to identify signaling components that contribute to D816V c-Kit-mediated transformation. SCF stimulates association of p85P13K with phosphorylated tyrosine 721 of wild-type c-Kit. Phosphatidylinositol 3 kinase (P13K) subsequently contributes to the activation of Akt and Jnks. In contrast, these studies demonstrated that the D816V c-Kit mutant was constitutively associated with phosphorylated p85 P13K, and, downstream of P13K, Jnk 1 and Jnk 2 were activated but Akt was not. Interestingly, Erks 1 and 2 were not constitutively activated by D816V c-Kit. Thus, D816V c-Kit maintains the activity of P13K but not of all signaling pathways activated by wild-type c-Kit. Further, all pathways downstream of P13K are not constitutively active in MIHC-D816V cells. Studies with a P13K inhibitor and D816V/Y721F c-Kit, a mutant incapable of recruiting P13K, indicate that constitutive activation of P13K through direct recruitment by D816V c-Kit plays a role in factor-independent growth of MIHC and is critical for tumorigenicity.

Original languageEnglish
Pages (from-to)1365-1373
Number of pages9
JournalBlood
Volume98
Issue number5
DOIs
Publication statusPublished - 1 Sep 2001

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Chian, R. J., Young, S., Danilkovitch-Miagkova, A., Rönnstrand, L., Leonard, E., Ferrao, P., ... Linnekin, D. (2001). Phosphatidylinositol 3 kinase contributes to the transformation of hematopoietic cells by the D816V c-Kit mutant. Blood, 98(5), 1365-1373. https://doi.org/10.1182/blood.V98.5.1365