Phospholipase C-gamma 1 and phosphatidylinositol 3 kinase are the downstream mediators of the PDGF receptor's mitogenic signal.
Upon ligand-induced tyrosine phosphorylation, the platelet-derived growth factor (PDGF) receptor (PDGFR) beta subunit associates with PLC-gamma 1, RasGAP, P13K, and a 64 kd protein. To determine the relative role of each of these associated proteins in PDGFR signaling, we constructed a PDGFR mutant (F5) unable to bind any of them and a panel of "add-back" mutants that could bind only one of the receptor-associated proteins. F5 PDGFR failed to activate PLC-gamma 1, P13K, or Ras and was unable to trigger DNA synthesis. Permitting association of F5 PDGFR with either PLC-gamma 1 or P13K restored Ras activation and a mitogenic response. Surprisingly, even though binding of the 64 kd protein almost fully restored Ras activation, it did not rescue the receptor's ability to trigger DNA synthesis. Thus Ras activation is insufficient to trigger PDGF-dependent DNA synthesis, and PLC-gamma 1 and P13K are independent downstream mediators of PDGF's mitogenic signal.
Valius
M
M
National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
Kazlauskas
A
A
D004247:DNA / Q000096:biosynthesis; D004252:DNA Mutational Analysis; D004789:Enzyme Activation; D020690:GTPase-Activating Proteins; D006801:Humans; D066298:In Vitro Techniques; D008938:Mitosis; D019869:Phosphatidylinositol 3-Kinases; D010770:Phosphotransferases / Q000502:physiology*; D019000:Phosphotyrosine; D010982:Platelet-Derived Growth Factor / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D017479:Receptors, Platelet-Derived Growth Factor / Q000502:physiology*; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000502:physiology*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D020703:ras GTPase-Activating Proteins
D020690:GTPase-Activating Proteins; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D020703:ras GTPase-Activating Proteins; D019000:Phosphotyrosine; D014443:Tyrosine; D004247:DNA; D010770:Phosphotransferases; D019869:Phosphatidylinositol 3-Kinases; D011505:Protein-Tyrosine Kinases; D017479:Receptors, Platelet-Derived Growth Factor; D010738:Type C Phospholipases