Donald Danforth Plant Science Center, 975 North Warson Road, St Louis, MO 63132, USA.
Department of Biology, Washington University, One Brookings Drive, Campus Box 1137, St Louis, MO 63130, USA.
New Phytol. 2011 Apr;190(1):35-48. doi: 10.1111/j.1469-8137.2010.03581.x. Epub 2010 Dec 22.
The repertoire of heterotrimeric G-proteins in plant species analyzed thus far is simple, with the presence of only two possible canonical heterotrimers in Arabidopsis and rice vs hundreds in animal systems. We assessed whether genome duplication events have resulted in the multiplicity of G-protein in plant species like soybean that would increase the complexity of G-protein networks. We identified and amplified four Gα, four Gβ and two Gγ proteins, analyzed their expression profile by quantitative PCR during different developmental stages. We purified the four Gα proteins and analyzed their guanosine-5'-triphosphate (GTP)-binding and GTPase activity. We performed yeast-based interaction analysis to assess the interaction specificity of different G-protein subunits. Our results show that all 10 G-protein genes are retained in the soybean genome and ubiquitously expressed. The four Gα proteins seem to be plasma membrane-localized. The G-protein genes have interesting expression profiles during seed development and germination. The four Gα proteins form two distinct groups based on their GTPase activity. Yeast-based interaction analyses predict that the proteins interact in most of the possible combinations, with some degree of interaction specificity between duplicated gene pairs. This research identifies the most elaborate heterotrimeric G-protein network known to date in the plant kingdom.
迄今为止,已分析的植物物种中的异三聚体 G 蛋白的种类很简单,拟南芥和水稻中仅有两种可能的典型异三聚体,而动物系统中则有数百种。我们评估了基因组复制事件是否导致像大豆这样的植物物种中 G 蛋白的多样性增加,从而增加 G 蛋白网络的复杂性。我们鉴定并扩增了四个 Gα、四个 Gβ和两个 Gγ蛋白,并通过定量 PCR 分析它们在不同发育阶段的表达谱。我们纯化了四个 Gα 蛋白,并分析了它们的鸟苷-5'-三磷酸 (GTP) 结合和 GTP 酶活性。我们进行了酵母相互作用分析,以评估不同 G 蛋白亚基的相互作用特异性。我们的研究结果表明,大豆基因组中保留了所有 10 个 G 蛋白基因,并广泛表达。这四个 Gα 蛋白似乎定位于质膜。G 蛋白基因在种子发育和萌发过程中具有有趣的表达模式。根据 GTPase 活性,四个 Gα 蛋白可分为两个不同的组。酵母相互作用分析预测,这些蛋白质在大多数可能的组合中相互作用,在重复基因对之间存在一定程度的相互作用特异性。这项研究确定了迄今为止在植物界中已知的最复杂的异三聚体 G 蛋白网络。