Department of Plant Biology, University of Minnesota, Saint Paul, Minnesota 55108, USA.
Plant Physiol. 2010 Apr;152(4):1889-900. doi: 10.1104/pp.109.150573. Epub 2010 Feb 10.
Sugars, such as sucrose and glucose, have been implicated in the regulation of diverse developmental events in plants and other organisms. We isolated an Arabidopsis (Arabidopsis thaliana) mutant, sugar-insensitive3 (sis3), that is resistant to the inhibitory effects of high concentrations of exogenous glucose and sucrose on early seedling development. In contrast to wild-type plants, sis3 mutants develop green, expanded cotyledons and true leaves when sown on medium containing high concentrations (e.g. 270 mm) of sucrose. Unlike some other sugar response mutants, sis3 exhibits wild-type responses to the inhibitory effects of abscisic acid and paclobutrazol, a gibberellic acid biosynthesis inhibitor, on seed germination. Map-based cloning revealed that SIS3 encodes a RING finger protein. Complementation of the sis3-2 mutant with a genomic SIS3 clone restored sugar sensitivity of sis3-2, confirming the identity of the SIS3 gene. Biochemical analyses demonstrated that SIS3 is functional in an in vitro ubiquitination assay and that the RING motif is sufficient for its activity. Our results indicate that SIS3 encodes a ubiquitin E3 ligase that is a positive regulator of sugar signaling during early seedling development.
糖,如蔗糖和葡萄糖,被认为参与了植物和其他生物体中多种发育事件的调节。我们分离出一个拟南芥(Arabidopsis thaliana)突变体,糖不敏感 3 型(sis3),它对高浓度外源葡萄糖和蔗糖对早期幼苗发育的抑制作用有抗性。与野生型植物相比,sis3 突变体在含有高浓度(例如 270mm)蔗糖的培养基上播种时,会发育出绿色、展开的子叶和真叶。与其他一些糖反应突变体不同,sis3 对脱落酸和赤霉素生物合成抑制剂多效唑对种子萌发的抑制作用表现出野生型反应。基于图谱的克隆表明,SIS3 编码一个 RING 指蛋白。用基因组 SIS3 克隆互补 sis3-2 突变体,恢复了 sis3-2 的糖敏感性,证实了 SIS3 基因的身份。生化分析表明,SIS3 在体外泛素化测定中具有功能,并且 RING 基序足以发挥其活性。我们的结果表明,SIS3 编码一种泛素 E3 连接酶,它是早期幼苗发育过程中糖信号的正调节剂。