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

被引:174
作者
Jones, N
Master, Z
Jones, J
Bouchard, D
Gunji, Y
Sasaki, H
Daly, R
Alitalo, K
Dumont, DJ
机构
[1] Sunnybrook & Womens Coll Hlth Sci Ctr, Div Canc Biol Res, Toronto, ON M4N 3M5, Canada
[2] Univ Toronto, Dept Med Biophys, Toronto, ON M4N 3M5, Canada
[3] Amgen Inst, Toronto, ON M5G 2G1, Canada
[4] Univ Helsinki, Haartman Inst, Mol Canc Biol Lab, FIN-00014 Helsinki, Finland
[5] Natl Canc Ctr, Res Inst, Chuo Ku, Tokyo 104, Japan
[6] St Vincents Hosp, Garvan Inst Med Res, Canc Res Program, Sydney, NSW 2010, Australia
关键词
D O I
10.1074/jbc.274.43.30896
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (Tyr(1100)). 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.
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页码:30896 / 30905
页数:10
相关论文
共 47 条
[1]   Intracellular signaling of the Ufo/Axl receptor tyrosine kinase is mediated mainly by a multi-substrate docking-site [J].
Braunger, J ;
Schleithoff, L ;
Schulz, AS ;
Kessler, H ;
Lammers, R ;
Ullrich, A ;
Bartram, CR ;
Janssen, JWG .
ONCOGENE, 1997, 14 (22) :2619-2631
[2]   THE 2-HYBRID SYSTEM - A METHOD TO IDENTIFY AND CLONE GENES FOR PROTEINS THAT INTERACT WITH A PROTEIN OF INTEREST [J].
CHIEN, CT ;
BARTEL, PL ;
STERNGLANZ, R ;
FIELDS, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (21) :9578-9582
[3]   INVIVO BINDING-PROPERTIES OF SH2 DOMAINS FROM GTPASE-ACTIVATING PROTEIN AND PHOSPHATIDYLINOSITOL 3-KINASE [J].
COOPER, JA ;
KASHISHIAN, A .
MOLECULAR AND CELLULAR BIOLOGY, 1993, 13 (03) :1737-1745
[4]   Isolation of Angiopoietin-1, a ligand for the TIE2 receptor, by secretion-trap expression cloning [J].
Davis, S ;
Aldrich, TH ;
Jones, PF ;
Acheson, A ;
Compton, DL ;
Jain, V ;
Ryan, TE ;
Bruno, J ;
Radziejewski, C ;
Maisonpierre, PC ;
Yancopoulos, GD .
CELL, 1996, 87 (07) :1161-1169
[5]   Mechanisms and consequences of activation of protein kinase B/Akt [J].
Downward, J .
CURRENT OPINION IN CELL BIOLOGY, 1998, 10 (02) :262-267
[6]  
DUMONT DJ, 1992, ONCOGENE, V7, P1471
[7]   Cardiovascular failure in mouse embryos deficient in VEGF receptor-3 [J].
Dumont, DJ ;
Jussila, L ;
Taipale, J ;
Lymboussaki, A ;
Mustonen, T ;
Pajusola, K ;
Breitman, M ;
Alitalo, K .
SCIENCE, 1998, 282 (5390) :946-949
[8]   DOMINANT-NEGATIVE AND TARGETED NULL MUTATIONS IN THE ENDOTHELIAL RECEPTOR TYROSINE KINASE, TEK, REVEAL A CRITICAL ROLE IN VASCULOGENESIS OF THE EMBRYO [J].
DUMONT, DJ ;
GRADWOHL, G ;
FONG, GH ;
PURI, MC ;
GERTSENSTEIN, M ;
AUERBACH, A ;
BREITMAN, ML .
GENES & DEVELOPMENT, 1994, 8 (16) :1897-1909
[9]   Analysis of Grb7 recruitment by heregulin-activated erbB receptors reveals a novel target selectivity for erbB3 [J].
Fiddes, RJ ;
Campbell, DH ;
Janes, PW ;
Sivertsen, SP ;
Sasaki, H ;
Wallasch, C ;
Daly, RJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (13) :7717-7724
[10]   A NOVEL GENETIC SYSTEM TO DETECT PROTEIN PROTEIN INTERACTIONS [J].
FIELDS, S ;
SONG, OK .
NATURE, 1989, 340 (6230) :245-246