Department of Chemistry, University of Washington , Seattle, Washington, U.S.A.
Department of Biochemistry, University of Washington , Seattle, Washington, U.S.A.
Small GTPases. 2020 Nov;11(6):413-420. doi: 10.1080/21541248.2018.1446697. Epub 2018 Apr 10.
RAS signaling pathways govern diverse cellular processes, are dynamic, and exhibit marked plasticity. Yet, these features also present a considerable obstacle to their study. Here, we report the use of a recently described RAS rheostat, Chemically Inducible Activator of RAS (CIAR), to study two poorly understood phenomena in RAS biology. First, we show that short-term activation of wild type endogenous RAS can desensitize cells to EGF stimulation. Second, we examine the phenomena of paradoxical activation of RAS/ERK signaling by RAF inhibitors. Specifically, we characterize the effects on RAS/ERK signaling kinetics of four RAF inhibitors, which stabilize distinct ATP-binding site conformations. These results demonstrate the utility of CIAR in conducting quantitative studies of complex features of RAS biology.
RAS 信号通路调控多种细胞过程，具有动态性和显著的可塑性。然而，这些特征也给它们的研究带来了相当大的障碍。在这里，我们报告了使用最近描述的 RAS 变阻器——化学诱导激活的 RAS（CIAR）来研究 RAS 生物学中两个尚未被充分理解的现象。首先，我们表明，野生型内源性 RAS 的短期激活可以使细胞对 EGF 刺激脱敏。其次，我们研究了 RAF 抑制剂引起的 RAS/ERK 信号悖论性激活的现象。具体来说，我们描述了四种 RAF 抑制剂对 RAS/ERK 信号动力学的影响，这四种抑制剂稳定了不同的 ATP 结合位点构象。这些结果证明了 CIAR 在对 RAS 生物学的复杂特征进行定量研究中的实用性。
