von Kriegsheim Alex, Baiocchi Daniela, Birtwistle Marc, Sumpton David, Bienvenut Willy, Morrice Nicholas, Yamada Kayo, Lamond Angus, Kalna Gabriella, Orton Richard, Gilbert David, Kolch Walter
Signalling and Proteomics Laboratory, The Beatson Institute for Cancer Research, Glasgow G61 1BD, UK.
MRC Protein Phosphorylation Unit, University of Dundee, Dundee DD1 5EH, Scotland, UK.
Nat Cell Biol. 2009 Dec;11(12):1458-64. doi: 10.1038/ncb1994. Epub 2009 Nov 22.
Extracellular signal-regulated kinase (ERK) controls fundamental cellular functions, including cell fate decisions. In PC12, cells shifting ERK activation from transient to sustained induces neuronal differentiation. As ERK associates with both regulators and effectors, we hypothesized that the mechanisms underlying the switch could be revealed by assessing the dynamic changes in ERK-interacting proteins that specifically occur under differentiation conditions. Using quantitative proteomics, we identified 284 ERK-interacting proteins. Upon induction of differentiation, 60 proteins changed their binding to ERK, including many proteins that were not known to participate in differentiation. We functionally characterized a subset, showing that they regulate the pathway at several levels and by different mechanisms, including signal duration, ERK localization, feedback, crosstalk with the Akt pathway and differential interaction and phosphorylation of transcription factors. Integrating these data with a mathematical model confirmed that ERK dynamics and differentiation are regulated by distributed control mechanisms rather than by a single master switch.
细胞外信号调节激酶（ERK）控制着包括细胞命运决定在内的基本细胞功能。在PC12细胞中，将ERK激活从短暂激活转变为持续激活会诱导神经元分化。由于ERK与调节因子和效应器都有关联，我们推测通过评估在分化条件下特异性发生的ERK相互作用蛋白的动态变化，可以揭示这种转变背后的机制。利用定量蛋白质组学，我们鉴定出了284种ERK相互作用蛋白。在诱导分化时，有60种蛋白质改变了它们与ERK的结合，其中包括许多此前未知参与分化过程的蛋白质。我们对其中一个子集进行了功能表征，结果表明它们在多个层面并通过不同机制调节该信号通路，包括信号持续时间、ERK定位、反馈、与Akt信号通路的串扰以及转录因子的差异相互作用和磷酸化。将这些数据与一个数学模型相结合证实，ERK动力学和分化是由分布式控制机制而非单一主开关调节的。
