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Identification of two juxtamembrane autophosphorylation sites in the PDGF beta-receptor; involvement in the interaction with Src family tyrosine kinases

Mori, Seijiro; Rönnstrand, Lars LU ; Yokote, Koutaro; Engström, Åke; Courtneidge, Sara A.; Claesson-Welsh, Lena and Heldin, Carl-Henrik (1993) In EMBO Journal 12(6). p.2257-2264
Abstract
Two novel sites of autophosphorylation were localized to the juxtamembrane segment of the human platelet-derived growth factor (PDGF) beta-receptor. To evaluate the importance of these phosphorylation sites, receptor mutants were made in which Tyr579, Tyr581 or both were replaced with phenylalanine residues; the receptor mutants were stably expressed in porcine aortic endothelial cells. Compared with the wild-type receptor, the Y579F and Y581F mutants were less able to mediate association with and activation of the Src family tyrosine kinases. The ability of these phosphorylation sites to mediate directly the binding of the Src family proteins was also demonstrated by using phosphotyrosine-containing synthetic peptides representing the... (More)
Two novel sites of autophosphorylation were localized to the juxtamembrane segment of the human platelet-derived growth factor (PDGF) beta-receptor. To evaluate the importance of these phosphorylation sites, receptor mutants were made in which Tyr579, Tyr581 or both were replaced with phenylalanine residues; the receptor mutants were stably expressed in porcine aortic endothelial cells. Compared with the wild-type receptor, the Y579F and Y581F mutants were less able to mediate association with and activation of the Src family tyrosine kinases. The ability of these phosphorylation sites to mediate directly the binding of the Src family proteins was also demonstrated by using phosphotyrosine-containing synthetic peptides representing the juxtamembrane sequence of the receptor. Both the Y579F and Y581F mutants were similar to the wild-type receptor with regard to their protein tyrosine kinase activity and ability to induce mitogenicity in response to PDGF-BB. A conclusive evaluation of the role of the Src family members in signal transduction could, however, not be made since our attempt to prevent completely the association by mutation of both Tyr579 and Tyr581, resulted in loss of kinase activity and was therefore not informative. The present data, together with previous observations, demonstrate a high degree of specificity in the interaction between different autophosphorylation sites in the PDGF beta-receptor and downstream components in the signal transduction pathway. (Less)
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Contribution to journal
publication status
published
subject
keywords
Amino Acid Sequence Base Sequence Binding Sites Cell Line Humans Molecular Sequence Data Oligodeoxyribonucleotides Phosphorylation Protein-Tyrosine Kinases/*metabolism Proto-Oncogene Proteins pp60(c-src)/*metabolism Receptors, Platelet-Derived Growth Factor/*metabolism Sequence Homology, Amino Acid
in
EMBO Journal
volume
12
issue
6
pages
2257 - 2264
publisher
Oxford University Press
ISSN
1460-2075
language
English
LU publication?
no
id
7be54f3e-bfbf-4874-96af-6121bdbd352e (old id 1784127)
alternative location
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC413454/
date added to LUP
2011-02-07 15:33:36
date last changed
2016-06-29 09:02:26
@misc{7be54f3e-bfbf-4874-96af-6121bdbd352e,
  abstract     = {Two novel sites of autophosphorylation were localized to the juxtamembrane segment of the human platelet-derived growth factor (PDGF) beta-receptor. To evaluate the importance of these phosphorylation sites, receptor mutants were made in which Tyr579, Tyr581 or both were replaced with phenylalanine residues; the receptor mutants were stably expressed in porcine aortic endothelial cells. Compared with the wild-type receptor, the Y579F and Y581F mutants were less able to mediate association with and activation of the Src family tyrosine kinases. The ability of these phosphorylation sites to mediate directly the binding of the Src family proteins was also demonstrated by using phosphotyrosine-containing synthetic peptides representing the juxtamembrane sequence of the receptor. Both the Y579F and Y581F mutants were similar to the wild-type receptor with regard to their protein tyrosine kinase activity and ability to induce mitogenicity in response to PDGF-BB. A conclusive evaluation of the role of the Src family members in signal transduction could, however, not be made since our attempt to prevent completely the association by mutation of both Tyr579 and Tyr581, resulted in loss of kinase activity and was therefore not informative. The present data, together with previous observations, demonstrate a high degree of specificity in the interaction between different autophosphorylation sites in the PDGF beta-receptor and downstream components in the signal transduction pathway.},
  author       = {Mori, Seijiro and Rönnstrand, Lars and Yokote, Koutaro and Engström, Åke and Courtneidge, Sara A. and Claesson-Welsh, Lena and Heldin, Carl-Henrik},
  issn         = {1460-2075},
  keyword      = {Amino Acid Sequence
Base Sequence
Binding Sites
Cell Line
Humans
Molecular Sequence Data
Oligodeoxyribonucleotides
Phosphorylation
Protein-Tyrosine Kinases/*metabolism
Proto-Oncogene Proteins pp60(c-src)/*metabolism
Receptors,Platelet-Derived Growth Factor/*metabolism
Sequence Homology,Amino Acid},
  language     = {eng},
  number       = {6},
  pages        = {2257--2264},
  publisher    = {ARRAY(0x79cece8)},
  series       = {EMBO Journal},
  title        = {Identification of two juxtamembrane autophosphorylation sites in the PDGF beta-receptor; involvement in the interaction with Src family tyrosine kinases},
  volume       = {12},
  year         = {1993},
}