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The activation loop tyrosine 823 is essential for the transforming capacity of the c-Kit oncogenic mutant D816V.

Agarwal, Shruti LU ; Kazi, Julhash U. LU ; Mohlin, Sofie LU ; Påhlman, Sven LU and Rönnstrand, Lars LU (2015) In Oncogene 34(35). p.4581-4590
Abstract
Oncogenic c-Kit mutations have been shown to display ligand-independent receptor activation and cell proliferation. A substitution of aspartate to valine at amino acid 816 (D816V) is one of the most commonly found oncogenic c-Kit mutations and is found in >90% of cases of mastocytosis and less commonly in germ-cell tumors, core-binding factor acute myeloid leukemia and mucosal melanomas. The mechanisms by which this mutation leads to constitutive activation and transformation are not fully understood. Previous studies have shown that the D816V mutation causes a structural change in the activation loop (A-loop), resulting in weaker binding of the A-loop to the juxtamembrane domain. In this paper, we have investigated the role of Y823,... (More)
Oncogenic c-Kit mutations have been shown to display ligand-independent receptor activation and cell proliferation. A substitution of aspartate to valine at amino acid 816 (D816V) is one of the most commonly found oncogenic c-Kit mutations and is found in >90% of cases of mastocytosis and less commonly in germ-cell tumors, core-binding factor acute myeloid leukemia and mucosal melanomas. The mechanisms by which this mutation leads to constitutive activation and transformation are not fully understood. Previous studies have shown that the D816V mutation causes a structural change in the activation loop (A-loop), resulting in weaker binding of the A-loop to the juxtamembrane domain. In this paper, we have investigated the role of Y823, the only tyrosine residue in the A-loop, and its role in oncogenic transformation by c-Kit/D816V by introducing the Y823F mutation. Although dispensable for the kinase activity of c-Kit/D816V, the presence of Y823 was crucial for cell proliferation and survival. Furthermore, mutation of Y823 selectively downregulates the Ras/Erk and Akt pathways as well as the phosphorylation of STAT5 and reduces the transforming capacity of the D816V/c-Kit in vitro. We further show that mice injected with cells expressing c-Kit/D816V/Y823F display significantly reduced tumor size as well as tumor weight compared with controls. Finally, microarray analysis, comparing Y823F/D816V cells with cells expressing c-Kit/D816V, demonstrate that mutation of Y823 causes upregulation of proapoptotic genes, whereas genes of survival pathways are downregulated. Thus, phosphorylation of Y823 is not necessary for kinase activation, but essential for the transforming ability of the c-Kit/D816V mutant.Oncogene advance online publication, 1 December 2014; doi:10.1038/onc.2014.383. (Less)
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author
organization
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type
Contribution to journal
publication status
published
subject
in
Oncogene
volume
34
issue
35
pages
4581 - 4590
publisher
Nature Publishing Group
external identifiers
  • pmid:25435369
  • wos:000360189300004
  • scopus:84940602883
ISSN
1476-5594
DOI
10.1038/onc.2014.383
project
CREATE Health
language
English
LU publication?
yes
id
9b0d007c-638d-47a3-8db5-1717c6f0caca (old id 4913654)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/25435369?dopt=Abstract
date added to LUP
2015-01-08 14:12:45
date last changed
2017-07-09 03:10:32
@article{9b0d007c-638d-47a3-8db5-1717c6f0caca,
  abstract     = {Oncogenic c-Kit mutations have been shown to display ligand-independent receptor activation and cell proliferation. A substitution of aspartate to valine at amino acid 816 (D816V) is one of the most commonly found oncogenic c-Kit mutations and is found in >90% of cases of mastocytosis and less commonly in germ-cell tumors, core-binding factor acute myeloid leukemia and mucosal melanomas. The mechanisms by which this mutation leads to constitutive activation and transformation are not fully understood. Previous studies have shown that the D816V mutation causes a structural change in the activation loop (A-loop), resulting in weaker binding of the A-loop to the juxtamembrane domain. In this paper, we have investigated the role of Y823, the only tyrosine residue in the A-loop, and its role in oncogenic transformation by c-Kit/D816V by introducing the Y823F mutation. Although dispensable for the kinase activity of c-Kit/D816V, the presence of Y823 was crucial for cell proliferation and survival. Furthermore, mutation of Y823 selectively downregulates the Ras/Erk and Akt pathways as well as the phosphorylation of STAT5 and reduces the transforming capacity of the D816V/c-Kit in vitro. We further show that mice injected with cells expressing c-Kit/D816V/Y823F display significantly reduced tumor size as well as tumor weight compared with controls. Finally, microarray analysis, comparing Y823F/D816V cells with cells expressing c-Kit/D816V, demonstrate that mutation of Y823 causes upregulation of proapoptotic genes, whereas genes of survival pathways are downregulated. Thus, phosphorylation of Y823 is not necessary for kinase activation, but essential for the transforming ability of the c-Kit/D816V mutant.Oncogene advance online publication, 1 December 2014; doi:10.1038/onc.2014.383.},
  author       = {Agarwal, Shruti and Kazi, Julhash U. and Mohlin, Sofie and Påhlman, Sven and Rönnstrand, Lars},
  issn         = {1476-5594},
  language     = {eng},
  number       = {35},
  pages        = {4581--4590},
  publisher    = {Nature Publishing Group},
  series       = {Oncogene},
  title        = {The activation loop tyrosine 823 is essential for the transforming capacity of the c-Kit oncogenic mutant D816V.},
  url          = {http://dx.doi.org/10.1038/onc.2014.383},
  volume       = {34},
  year         = {2015},
}