Analysis of Ras/ERK Compartmentalization by Subcellular Fractionation
Analysis of Ras/ERK Compartmentalization by Subcellular Fractionation
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DOI：
10.1007/978-1-4939-6424-6_11
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发表时间：
2017-01-01
期刊：
影响因子：
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通讯作者：
Casar, Berta
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文献类型：
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作者：
Agudo-Ibanez, Lorena;Crespo, Piero;Casar, Berta
大量的刺激使用Ras/Raf/MEK/ERK信号级联从其同源受体传递信号，以调节多种细胞功能，包括关键过程，如增殖、细胞周期进展、分化和存活。反应的持续时间、强度和特异性部分地由信号传导中介体的区室化/亚细胞定位控制。Ras蛋白存在于不同的质膜微区和内膜中。在这些定位中，Ras受到位点特异性调节机制的影响，独特地参与效应子途径并开启不同的遗传程序以产生多种生物反应。Ras效应子途径导致ERK激活也受到空间相关的调控过程。大约一半的ERK 1/2底物存在于细胞核中，主要作为转录因子发挥作用。另一半存在于细胞质和其他细胞器中。这种亚细胞分布增强了Ras/ERK级联的复杂性，并构成了赋予其信号可变性的重要机制，这使得它们能够参与多种功能的调节。因此，分析Ras/ERK级联成员的亚细胞区室化构成了研究Ras/ERK信号诱发的特定生物反应时要考虑的重要因素。在此，我们描述了用于这种目的的方法。
A vast number of stimuli use the Ras/Raf/MEK/ERK signaling cascade to transmit signals from their cognate receptors, in order to regulate multiple cellular functions, including key processes such as proliferation, cell cycle progression, differentiation, and survival. The duration, intensity and specificity of the responses are, in part, controlled by the compartmentalization/subcellular localization of the signaling intermediaries. Ras proteins are found in different plasma membrane microdomains and endomembranes. At these localizations, Ras is subject to site-specific regulatory mechanisms, distinctively engaging effector pathways and switching-on diverse genetic programs to generate a multitude of biological responses. The Ras effector pathway leading to ERKs activation is also subject to space-related regulatory processes. About half of ERK1/2 substrates are found in the nucleus and function mainly as transcription factors. The other half resides in the cytosol and other cellular organelles. Such subcellular distribution enhances the complexity of the Ras/ERK cascade and constitutes an essential mechanism to endow variability to its signals, which enables their participation in the regulation of a broad variety of functions. Thus, analyzing the subcellular compartmentalization of the members of the Ras/ERK cascade constitutes an important factor to be taken into account when studying specific biological responses evoked by Ras/ERK signals. Herein, we describe methods for such purpose.