A role for Src in signal relay by the platelet-derived growth factor alpha receptor.
Previous studies have shown that Src is required for platelet-derived growth factor (PDGF)-dependent cell cycle progression in fibroblasts. Since fibroblasts usually express both PDGF receptors (PDGFRs), these findings suggested that Src was mandatory for signal relay by both the alpha and betaPDGFRs. In this study, we have focused on the role of Src in signal relay by the alphaPDGFR. In response to stimulation with PDGF-AA, which selectively engages the alphaPDGFR, Src family members (Src) associated with the alphaPDGFR and Src kinase were activated. A mutant receptor, in which tyrosines 572 and 574 were replaced with phenylalanine (F72/74), failed to efficiently associate with Src or activate Src. The wild type (WT) and F72/74 receptors induced the expression of c-myc and c-fos to comparable levels. Furthermore, an equivalent extent of PDGF-dependent soft agar growth was observed in cells expressing the WT or the F72/74 alphaPDGFR. Comparing the ability of these two receptors to initiate tyrosine phosphorylation of signaling molecules indicated that both receptors mediated phosphorylation of the receptor itself, phospholipase Cgamma 1, and SHP-2 to similar levels. In contrast, the F72/74 receptor triggered phosphorylation of Shc to 1 and 20% of the WT levels for the 55- and 46-kDa Shc isoforms, respectively. These findings indicate that after exposure of cells to PDGF-AA, Src stably associates with the alphaPDGFR, and Src activity is increased. Furthermore, Src is required for the PDGF-dependent phosphorylation of signaling molecules such as Shc. Finally, activation of Src during the G0/G1 transition does not appear to be required for latter cell cycle events such as induction of c-myc or cell proliferation.
Gelderloos
J A
JA
The Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.
Rosenkranz
S
S
Bazenet
C
C
Kazlauskas
A
A
D016475:3T3 Cells; D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000235:genetics; D004789:Enzyme Activation; D005786:Gene Expression Regulation / Q000235:genetics; D051379:Mice; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000032:analysis; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D017479:Receptors, Platelet-Derived Growth Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D019061:src-Family Kinases / Q000378:metabolism*
D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; C075278:platelet-derived growth factor A; D019000:Phosphotyrosine; D017479:Receptors, Platelet-Derived Growth Factor; D019061:src-Family Kinases