Tyr-716 in the platelet-derived growth factor beta-receptor kinase insertis involved in GRB2 binding and Ras activation

Arvidsson, Ann-KristinUppsala universitet,Ludwiginstitutet för cancerforskning,Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden (författare)

Ligand stimulation of the platelet-derived growth factor (PDGF) beta-receptor leads to activation of its intrinsic tyrosine kinase and autophosphorylation of the intracellular part of the receptor. The autophosphorylated tyrosine residues mediate interactions with downstream signal transduction molecules and thereby initiate different signalling pathways. A pathway leading to activation of the GTP-binding protein Ras involves the adaptor molecule GRB2. Here we show that Tyr-716, a novel autophosphorylation site in the PDGF beta-receptor kinase insert, mediates direct binding of GRB2 in vitro and in vivo. In a panel of mutant PDGF beta-receptors, in which Tyr-716 and the previously known autophosphorylation sites were individually mutated, only PDGFR beta Y716F failed to bind GRB2. Furthermore, a synthetic phosphorylated peptide containing Tyr-716 bound GRB2, and this peptide specifically interrupted the interaction between GRB2 and the wild-type receptor. In addition, the Y716(P) peptide significantly decreased the amount of GTP bound to Ras in response to PDGF in permeabilized fibroblasts as well as in porcine aortic endothelial cells expressing transfected PDGF beta-receptors. The mutant PDGFR beta Y716F still mediated activation of mitogen-activated protein kinases and an increased DNA synthesis in response to PDGF, indicating that multiple signal transduction pathways transduce mitogenic signals from the activated PDGF beta-receptor.

Rupp, EvaUppsala universitet,Ludwiginstitutet för cancerforskning,Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden (författare)
Nånberg, Eewa,1957-Uppsala universitet,Institutionen för genetik och patologi,Department of Pathology, University Hospital, Uppsala, Sweden,Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi(Swepub:kau)eewanan (författare)
Downward, JulianSignal Transduction Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom (författare)
Rönnstrand, LarsUppsala universitet,Ludwiginstitutet för cancerforskning,Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden (författare)
Wennström, StefanUppsala universitet,Ludwiginstitutet för cancerforskning,Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden (författare)
Schlessinger, JosephDepartment of Pharmacology, New York University Medical Center, New York New York 10016, USA (författare)
Heldin, Carl-HenrikUppsala universitet,Ludwiginstitutet för cancerforskning,Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden (författare)
Claesson-Welsh, LenaUppsala universitet,Ludwiginstitutet för cancerforskning,Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden (författare)
Uppsala universitetLudwiginstitutet för cancerforskning (creator_code:org_t)

Ingår i:Molecular and Cellular Biology: American Society for Microbiology14:10, s. 6715-67260270-73061098-5549

Av författaren/redakt...: Arvidsson, Ann-K ...; Rupp, Eva; Nånberg, Eewa, 1 ...; Downward, Julian; Rönnstrand, Lars; Wennström, Stefa ...; visa fler...; Schlessinger, Jo ...; Heldin, Carl-Hen ...; Claesson-Welsh, ...; visa färre...

Om ämnet

Av lärosätet: Uppsala universitet; Örebro universitet; Karlstads universitet

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