Bakal Chris, Linding Rune, Llense Flora, Heffern Elleard, Martin-Blanco Enrique, Pawson Tony, Perrimon Norbert
Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02215, USA.
Science. 2008 Oct 17;322(5900):453-6. doi: 10.1126/science.1158739.
Cellular signaling networks have evolved to enable swift and accurate responses, even in the face of genetic or environmental perturbation. Thus, genetic screens may not identify all the genes that regulate different biological processes. Moreover, although classical screening approaches have succeeded in providing parts lists of the essential components of signaling networks, they typically do not provide much insight into the hierarchical and functional relations that exist among these components. We describe a high-throughput screen in which we used RNA interference to systematically inhibit two genes simultaneously in 17,724 combinations to identify regulators of Drosophila JUN NH(2)-terminal kinase (JNK). Using both genetic and phosphoproteomics data, we then implemented an integrative network algorithm to construct a JNK phosphorylation network, which provides structural and mechanistic insights into the systems architecture of JNK signaling.
细胞信号网络已经进化到能够实现快速而准确的反应，即使面对基因或环境扰动也是如此。因此，基因筛选可能无法识别所有调节不同生物过程的基因。此外，尽管经典筛选方法成功地提供了信号网络基本组成部分的清单，但它们通常无法深入了解这些组成部分之间存在的层次和功能关系。我们描述了一种高通量筛选方法，其中我们使用RNA干扰以17724种组合同时系统地抑制两个基因，以鉴定果蝇JUN氨基末端激酶（JNK）的调节因子。然后，我们利用遗传和磷酸化蛋白质组学数据，实施了一种整合网络算法来构建JNK磷酸化网络，该网络为JNK信号传导的系统架构提供了结构和机制方面的见解。
