Swain Stephen M, Tseng Tong-Seung, Thornton Tina M, Gopalraj Manjula, Olszewski Neil E
Department of Plant Biology and Plant Molecular Genetics Institute, University of Minnesota, St. Paul, MN 55108, USA.
Plant Physiol. 2002 Jun;129(2):605-15. doi: 10.1104/pp.020002.
SPY (SPINDLY) encodes a putative O-linked N-acetyl-glucosamine transferase that is genetically defined as a negatively acting component of the gibberellin (GA) signal transduction pathway. Analysis of Arabidopsis plants containing a SPY::GUS reporter gene reveals that SPY is expressed throughout the life of the plant and in most plant organs examined. In addition to being expressed in all organs where phenotypes due to spy mutations have been reported, SPY::GUS is expressed in the root. Examination of the roots of wild-type, spy, and gai plants revealed phenotypes indicating that SPY and GAI play a role in root development. A second SPY::GUS reporter gene lacking part of the SPY promoter was inactive, suggesting that sequences in the first exon and/or intron are required for detectable expression. Using both subcellular fractionation and visualization of a SPY-green fluorescent protein fusion protein that is able to rescue the spy mutant phenotype, the majority of SPY protein was shown to be present in the nucleus. This result is consistent with the nuclear localization of other components of the GA response pathway and suggests that SPY's role as a negative regulator of GA signaling involves interaction with other nuclear proteins and/or O-N-acetyl-glucosamine modification of these proteins.
SPY(细长)编码一种假定的O-连接的N-乙酰葡糖胺转移酶,在遗传学上被定义为赤霉素(GA)信号转导途径的负向作用成分。对含有SPY::GUS报告基因的拟南芥植株的分析表明,SPY在植物的整个生命周期以及所检测的大多数植物器官中均有表达。除了在已报道因spy突变而出现表型的所有器官中表达外,SPY::GUS在根中也有表达。对野生型、spy和gai植株的根进行检查,发现了一些表型,表明SPY和GAI在根的发育中发挥作用。第二个缺少部分SPY启动子的SPY::GUS报告基因没有活性,这表明第一个外显子和/或内含子中的序列是可检测表达所必需的。通过亚细胞分级分离以及对能够挽救spy突变体表型的SPY-绿色荧光蛋白融合蛋白的可视化分析,发现大多数SPY蛋白存在于细胞核中。这一结果与GA反应途径其他成分的核定位一致,并表明SPY作为GA信号的负调节因子,其作用涉及与其他核蛋白相互作用和/或对这些蛋白进行O-N-乙酰葡糖胺修饰。