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Identification of Y589 and Y599 in the juxamembrane domain of Flt3 as ligand-induced autophosphorylation sites involved in binding of Src family kinases and the protein tyrosine phosphatase SHP2

Heiss, Elke LU ; Masson, Kristina LU ; Sundberg, Christina LU ; Pedersen, Malin LU ; Sun, Jianmin LU ; Bengtsson, Susanne LU and Rönnstrand, Lars LU orcid (2006) In Blood 108(5). p.1542-1550
Abstract
Early signal relay steps upon ligand binding to the receptor tyrosine kinase Flt3, i.e. sites of Flt3 autophosphorylation and subsequent docking partners, are mainly unresolved. By immunoprecipitation of specific tryptic peptides contained in the juxtamembrane region

of human Flt3 and subsequent radiosequencing we identified the tyrosine residues 572, 589, 591 and 599 as in vivo autophosphorylation sites. Focusing on Y589 and Y599, we

examined Flt3-ligand-mediated responses in WT-Flt3-, Y589F-Flt3- and Y599F-Flt3-expressing 32D cells. Compared to WT-Flt3-32D cells upon ligand-stimulation, 32DY589F-

Flt3 showed enhanced Erk activation and proliferation/survival whereas 32DY599F-Flt3 cells hereby displayed... (More)
Early signal relay steps upon ligand binding to the receptor tyrosine kinase Flt3, i.e. sites of Flt3 autophosphorylation and subsequent docking partners, are mainly unresolved. By immunoprecipitation of specific tryptic peptides contained in the juxtamembrane region

of human Flt3 and subsequent radiosequencing we identified the tyrosine residues 572, 589, 591 and 599 as in vivo autophosphorylation sites. Focusing on Y589 and Y599, we

examined Flt3-ligand-mediated responses in WT-Flt3-, Y589F-Flt3- and Y599F-Flt3-expressing 32D cells. Compared to WT-Flt3-32D cells upon ligand-stimulation, 32DY589F-

Flt3 showed enhanced Erk activation and proliferation/survival whereas 32DY599F-Flt3 cells hereby displayed substantially diminished responses. Both pY589 and pY599 were identified as association sites for signal relay molecules including Src family kinases and SHP2. Consistently, 32D-Y589F-Flt3 and 32D-Y599F-Flt3 showed

decreased FL-triggered activation of Src family kinases. Interference with the Srcdependent negative regulation of Flt3 signaling may account for the enhanced mitogenic

response of Y589F-Flt3. Y599 was additionally found to interact with the protein tyrosine phosphatase SHP2 in a phosphorylation dependent manner. As Y599F-Flt3-32D

was unable to associate with and to phosporylate SHP2 and since silencing of SHP2 in WT-Flt3-expressing cells mimicked the Y599F-Flt3-phenotype we hypothesize that

recruitment of SHP2 to pY599 contributes to FL-mediated Erk activation and proliferation. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Blood
volume
108
issue
5
pages
1542 - 1550
publisher
American Society of Hematology
external identifiers
  • wos:000240271500024
  • pmid:16684964
  • scopus:33748192963
ISSN
1528-0020
DOI
10.1182/blood-2005-07-008896
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Experimental Clinical Chemistry (013016010)
id
62204146-a2ae-4d73-836b-a89128bdcc43 (old id 1137451)
date added to LUP
2016-04-01 12:32:54
date last changed
2021-08-11 05:21:59
@article{62204146-a2ae-4d73-836b-a89128bdcc43,
  abstract     = {Early signal relay steps upon ligand binding to the receptor tyrosine kinase Flt3, i.e. sites of Flt3 autophosphorylation and subsequent docking partners, are mainly unresolved. By immunoprecipitation of specific tryptic peptides contained in the juxtamembrane region<br/><br>
of human Flt3 and subsequent radiosequencing we identified the tyrosine residues 572, 589, 591 and 599 as in vivo autophosphorylation sites. Focusing on Y589 and Y599, we<br/><br>
examined Flt3-ligand-mediated responses in WT-Flt3-, Y589F-Flt3- and Y599F-Flt3-expressing 32D cells. Compared to WT-Flt3-32D cells upon ligand-stimulation, 32DY589F-<br/><br>
Flt3 showed enhanced Erk activation and proliferation/survival whereas 32DY599F-Flt3 cells hereby displayed substantially diminished responses. Both pY589 and pY599 were identified as association sites for signal relay molecules including Src family kinases and SHP2. Consistently, 32D-Y589F-Flt3 and 32D-Y599F-Flt3 showed<br/><br>
decreased FL-triggered activation of Src family kinases. Interference with the Srcdependent negative regulation of Flt3 signaling may account for the enhanced mitogenic<br/><br>
response of Y589F-Flt3. Y599 was additionally found to interact with the protein tyrosine phosphatase SHP2 in a phosphorylation dependent manner. As Y599F-Flt3-32D<br/><br>
was unable to associate with and to phosporylate SHP2 and since silencing of SHP2 in WT-Flt3-expressing cells mimicked the Y599F-Flt3-phenotype we hypothesize that<br/><br>
recruitment of SHP2 to pY599 contributes to FL-mediated Erk activation and proliferation.},
  author       = {Heiss, Elke and Masson, Kristina and Sundberg, Christina and Pedersen, Malin and Sun, Jianmin and Bengtsson, Susanne and Rönnstrand, Lars},
  issn         = {1528-0020},
  language     = {eng},
  number       = {5},
  pages        = {1542--1550},
  publisher    = {American Society of Hematology},
  series       = {Blood},
  title        = {Identification of Y589 and Y599 in the juxamembrane domain of Flt3 as ligand-induced autophosphorylation sites involved in binding of Src family kinases and the protein tyrosine phosphatase SHP2},
  url          = {http://dx.doi.org/10.1182/blood-2005-07-008896},
  doi          = {10.1182/blood-2005-07-008896},
  volume       = {108},
  year         = {2006},
}