Border Control-A Membrane-Linked Interactome of Arabidopsis
Border Control-A Membrane-Linked Interactome of Arabidopsis
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DOI：
10.1126/science.1251358
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发表时间：
2014-05-16
期刊：
影响因子：
56.9
通讯作者：
Frommer, Wolf B.
中科院分区：
文献类型：
--
作者：
Jones, Alexander M.;Xuan, Yuanhu;Frommer, Wolf B.
细胞膜作为信号平台，控制着溶质的运输。膜受体、转运体和酶通过蛋白质-蛋白质相互作用与细胞内过程进行通讯。利用覆盖6.4×10(6)对测试空间的裂解泛素酵母双杂交筛选，我们从拟南芥中鉴定了12,102个膜/信号蛋白相互作用。除了证实了预期的相互作用，如异三聚体G蛋白亚基相互作用和水通道蛋白齐聚，99%的相互作用以前是未知的。在植物裂解-绿色荧光蛋白相互作用实验中，相互作用的正交率为32%，这与文献中收集的已知相互作用的确认率(38%)在统计学上没有区别。膜蛋白运输、周转和磷酸化的调节关系包括通过脱落酸信号调节钾通道活性，通过WNK激酶调节转运体活性，以及通过与运输相关的蛋白调节油菜素内酯受体激酶。这些例子强调了膜/信号蛋白相互作用网络在植物和其他生物体的基因发现和假说产生中的作用。
Cellular membranes act as signaling platforms and control solute transport. Membrane receptors, transporters, and enzymes communicate with intracellular processes through protein-protein interactions. Using a split-ubiquitin yeast two-hybrid screen that covers a test-space of 6.4 x 10(6) pairs, we identified 12,102 membrane/signaling protein interactions from Arabidopsis. Besides confirmation of expected interactions such as heterotrimeric G protein subunit interactions and aquaporin oligomerization, >99% of the interactions were previously unknown. Interactions were confirmed at a rate of 32% in orthogonal in planta split-green flourescent protein interaction assays, which was statistically indistinguishable from the confirmation rate for known interactions collected from literature (38%). Regulatory associations in membrane protein trafficking, turnover, and phosphorylation include regulation of potassium channel activity through abscisic acid signaling, transporter activity by a WNK kinase, and a brassinolide receptor kinase by trafficking-related proteins. These examples underscore the utility of the membrane/signaling protein interaction network for gene discovery and hypothesis generation in plants and other organisms.