DeMali K A, Kazlauskas A
Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.
Mol Cell Biol. 1998 Apr;18(4):2014-22. doi: 10.1128/MCB.18.4.2014.
The basal activity of Src family kinases is readily detectable throughout the cell cycle and increases by two- to fivefold upon acute stimulation of cells with growth factors such as platelet-derived growth factor. Previous reports have demonstrated a requirement for Src activity for the G1/S and G2/M transitions. With a chimeric alpha-beta PDGF receptor (PDGFR) expressed in fibroblasts, we have investigated the importance of the PDGF-mediated increase in Src activity at the G0/G1 transition for subsequent cell cycle events. A mutant PDGFR chimera that was not able to detectably associate with or activate Src was compromised in its ability to mediate tyrosine phosphorylation of receptor-associated signaling molecules and initiated a submaximal activation of Erk. In contrast to these early cell cycle events, later responses such as entry of cells into S phase and cell proliferation proceeded normally when Src activity did not increase following acute stimulation with PDGF. We conclude that the initial burst of Src activity is required for efficient tyrosine phosphorylation of receptor-associated proteins such as PLCgamma, RasGAP, Shc, and SHP-2 and for maximal activation of Erk. Surprisingly, these events are not required for PDGF-dependent cell proliferation. Finally, later cell cycle events do not require that Src be activated at the G0/G1 transition and leave open the possibility that events such as the G1/S transition require the basal Src activity and/or activation of Src at later times in G1.
Src家族激酶的基础活性在整个细胞周期中都很容易检测到,在用血小板衍生生长因子等生长因子急性刺激细胞后,其活性会增加两到五倍。先前的报道表明,Src活性对于G1/S和G2/M期转换是必需的。通过在成纤维细胞中表达嵌合的α-β血小板衍生生长因子受体(PDGFR),我们研究了PDGF介导的在G0/G1期转换时Src活性增加对于后续细胞周期事件的重要性。一种无法与Src可检测地结合或激活Src的突变型PDGFR嵌合体,在介导受体相关信号分子的酪氨酸磷酸化以及启动Erk的次最大激活方面存在缺陷。与这些早期细胞周期事件相反,当用PDGF急性刺激后Src活性没有增加时,诸如细胞进入S期和细胞增殖等后期反应仍正常进行。我们得出结论,Src活性的初始爆发对于受体相关蛋白如PLCγ、RasGAP、Shc和SHP-2的有效酪氨酸磷酸化以及Erk的最大激活是必需的。令人惊讶的是,这些事件对于PDGF依赖性细胞增殖并非必需。最后,后期细胞周期事件并不要求Src在G0/G1期转换时被激活,这使得诸如G1/S期转换等事件可能需要基础Src活性和/或在G1期后期激活Src成为可能。