Membrane ruffling and chemotaxis transduced by the PDGF beta-receptor require the binding site for phosphatidylinositol 3' kinase.
Activation of the platelet-derived growth factor (PDGF) beta-receptor results in motility responses in the forms of membrane ruffling and chemotaxis. Porcine aortic endothelial cells expressing the PDGF beta-receptor or a chimeric fibroblast growth factor (FGF) receptor, in which the endogenous kinase insert was replaced with the corresponding region from the PDGF beta-receptor, migrated efficiently towards a concentration gradient of PDGF-BB and bFGF, respectively, and exhibited both pronounced edge ruffling and circular membrane ruffling in response to ligand-stimulation. The wildtype FGF receptor-1 showed weak or no response in these assays. Further analyses were conducted on mutant receptors, in which tyrosine residues that can serve as autophosphorylation sites and thereby mediate interactions with specific signal transduction molecules, were changed to phenylalanine residues. Each one of the analysed mutants were mitogenically active, however, a mutant in which Tyr740 and Tyr751 were replaced failed to mediate ruffling and chemotaxis. These two residues are implicated in the binding of phosphatidylinositol 3' kinase. The notion that this enzyme is involved in PDGF beta-receptor-induced cell motility is furthermore supported by the finding that another mutant, in which Met743 and Met754 were replaced, and which failed to interact with phosphatidylinositol 3' kinase, was also unable to mediate motility responses.
Wennström
S
S
Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden.
Siegbahn
A
A
Yokote
K
K
Arvidsson
A K
AK
Heldin
C H
CH
Mori
S
S
Claesson-Welsh
L
L
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002465:Cell Movement / Q000502:physiology; D002633:Chemotaxis / Q000502:physiology*; D004247:DNA / Q000378:metabolism; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D009154:Mutation; D019869:Phosphatidylinositol 3-Kinases; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism* / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D017479:Receptors, Platelet-Derived Growth Factor / Q000032:analysis / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D013552:Swine; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D014316:Tritium; D004247:DNA; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D011505:Protein-Tyrosine Kinases; D017479:Receptors, Platelet-Derived Growth Factor; D013936:Thymidine