Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration

Nina Jones, Zubin Master, Jamie Jones, Denis Bouchard, Yuji Gunji, Hiroki Sasaki, Roger Daly, Kari Alitalo, Daniel J. Dumont

Research output: Contribution to journalArticle

171 Citations (Scopus)

Abstract

The Tek/Tie2 receptor tyrosine kinase plays a pivotal role in vascular and hematopoietic development. To study the signal transduction pathways that are mediated by this receptor, we have used the yeast two-hybrid system to identify signaling molecules that associate with the phosphorylated Tek receptor. Using this approach, we demonstrate that five molecules, Grb2, Grb7, Grb14, Shp2, and the p85 subunit of phosphatidylinositol 3-kinase can interact with Tek in a phosphotyrosine-dependent manner through their SH2 domains. Mapping of the binding sites of these molecules on Tek reveals the presence of a multisubstrate docking site in the carboxyl tail of Tek (Tyr1100). Mutation of this site abrogates binding of Grb2 and Grb7 to Tek in vivo, and this site is required for tyrosine phosphorylation of Grb7 and p85 in vivo. Furthermore, stimulation of Tek-expressing cells with Angiopoietin-1 results in phosphorylation of both Tek and p85 and in activation of endothelial cell migration and survival pathways that are dependent in part on phosphatidylinositol 3-kinase. Taken together, these results demonstrate that Angiopoietin-1-induced signaling from the Tek receptor is mediated by a multifunctional docking site that is responsible for activation of both cell migration and cell survival pathways.

Original languageEnglish (US)
Pages (from-to)30896-30905
Number of pages10
JournalJournal of Biological Chemistry
Volume274
Issue number43
DOIs
StatePublished - Oct 22 1999
Externally publishedYes

Fingerprint

TIE-2 Receptor
Phosphatidylinositol 3-Kinase
Angiopoietin-1
Cell Movement
Cell Survival
Phosphorylation
Cells
Molecules
Chemical activation
Binding Sites
Signal transduction
Phosphotyrosine
Endothelial cells
Hybrid systems
Yeast
Protein-Tyrosine Kinases
Two-Hybrid System Techniques
src Homology Domains
Tyrosine
Blood Vessels

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration. / Jones, Nina; Master, Zubin; Jones, Jamie; Bouchard, Denis; Gunji, Yuji; Sasaki, Hiroki; Daly, Roger; Alitalo, Kari; Dumont, Daniel J.

In: Journal of Biological Chemistry, Vol. 274, No. 43, 22.10.1999, p. 30896-30905.

Research output: Contribution to journalArticle

Jones, Nina ; Master, Zubin ; Jones, Jamie ; Bouchard, Denis ; Gunji, Yuji ; Sasaki, Hiroki ; Daly, Roger ; Alitalo, Kari ; Dumont, Daniel J. / Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 43. pp. 30896-30905.
@article{222eb186f2d04ceb9f47b96ffaaaa2de,
title = "Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration",
abstract = "The Tek/Tie2 receptor tyrosine kinase plays a pivotal role in vascular and hematopoietic development. To study the signal transduction pathways that are mediated by this receptor, we have used the yeast two-hybrid system to identify signaling molecules that associate with the phosphorylated Tek receptor. Using this approach, we demonstrate that five molecules, Grb2, Grb7, Grb14, Shp2, and the p85 subunit of phosphatidylinositol 3-kinase can interact with Tek in a phosphotyrosine-dependent manner through their SH2 domains. Mapping of the binding sites of these molecules on Tek reveals the presence of a multisubstrate docking site in the carboxyl tail of Tek (Tyr1100). Mutation of this site abrogates binding of Grb2 and Grb7 to Tek in vivo, and this site is required for tyrosine phosphorylation of Grb7 and p85 in vivo. Furthermore, stimulation of Tek-expressing cells with Angiopoietin-1 results in phosphorylation of both Tek and p85 and in activation of endothelial cell migration and survival pathways that are dependent in part on phosphatidylinositol 3-kinase. Taken together, these results demonstrate that Angiopoietin-1-induced signaling from the Tek receptor is mediated by a multifunctional docking site that is responsible for activation of both cell migration and cell survival pathways.",
author = "Nina Jones and Zubin Master and Jamie Jones and Denis Bouchard and Yuji Gunji and Hiroki Sasaki and Roger Daly and Kari Alitalo and Dumont, {Daniel J.}",
year = "1999",
month = "10",
day = "22",
doi = "10.1074/jbc.274.43.30896",
language = "English (US)",
volume = "274",
pages = "30896--30905",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "43",

}

TY - JOUR

T1 - Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration

AU - Jones, Nina

AU - Master, Zubin

AU - Jones, Jamie

AU - Bouchard, Denis

AU - Gunji, Yuji

AU - Sasaki, Hiroki

AU - Daly, Roger

AU - Alitalo, Kari

AU - Dumont, Daniel J.

PY - 1999/10/22

Y1 - 1999/10/22

N2 - The Tek/Tie2 receptor tyrosine kinase plays a pivotal role in vascular and hematopoietic development. To study the signal transduction pathways that are mediated by this receptor, we have used the yeast two-hybrid system to identify signaling molecules that associate with the phosphorylated Tek receptor. Using this approach, we demonstrate that five molecules, Grb2, Grb7, Grb14, Shp2, and the p85 subunit of phosphatidylinositol 3-kinase can interact with Tek in a phosphotyrosine-dependent manner through their SH2 domains. Mapping of the binding sites of these molecules on Tek reveals the presence of a multisubstrate docking site in the carboxyl tail of Tek (Tyr1100). Mutation of this site abrogates binding of Grb2 and Grb7 to Tek in vivo, and this site is required for tyrosine phosphorylation of Grb7 and p85 in vivo. Furthermore, stimulation of Tek-expressing cells with Angiopoietin-1 results in phosphorylation of both Tek and p85 and in activation of endothelial cell migration and survival pathways that are dependent in part on phosphatidylinositol 3-kinase. Taken together, these results demonstrate that Angiopoietin-1-induced signaling from the Tek receptor is mediated by a multifunctional docking site that is responsible for activation of both cell migration and cell survival pathways.

AB - The Tek/Tie2 receptor tyrosine kinase plays a pivotal role in vascular and hematopoietic development. To study the signal transduction pathways that are mediated by this receptor, we have used the yeast two-hybrid system to identify signaling molecules that associate with the phosphorylated Tek receptor. Using this approach, we demonstrate that five molecules, Grb2, Grb7, Grb14, Shp2, and the p85 subunit of phosphatidylinositol 3-kinase can interact with Tek in a phosphotyrosine-dependent manner through their SH2 domains. Mapping of the binding sites of these molecules on Tek reveals the presence of a multisubstrate docking site in the carboxyl tail of Tek (Tyr1100). Mutation of this site abrogates binding of Grb2 and Grb7 to Tek in vivo, and this site is required for tyrosine phosphorylation of Grb7 and p85 in vivo. Furthermore, stimulation of Tek-expressing cells with Angiopoietin-1 results in phosphorylation of both Tek and p85 and in activation of endothelial cell migration and survival pathways that are dependent in part on phosphatidylinositol 3-kinase. Taken together, these results demonstrate that Angiopoietin-1-induced signaling from the Tek receptor is mediated by a multifunctional docking site that is responsible for activation of both cell migration and cell survival pathways.

UR - http://www.scopus.com/inward/record.url?scp=0032695794&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032695794&partnerID=8YFLogxK

U2 - 10.1074/jbc.274.43.30896

DO - 10.1074/jbc.274.43.30896

M3 - Article

VL - 274

SP - 30896

EP - 30905

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 43

ER -