Curaba Julien, Herzog Michel, Vachon Gilles
Laboratoire de Génétique Moléculaire des Plantes, CNRS UMR 5575, Université Joseph Fourier, CERMO B.P. 53, F-38041 Grenoble Cedex 9, France.
Plant J. 2003 Jan;33(2):305-17. doi: 10.1046/j.1365-313x.2003.01622.x.
Trichomes of Arabidopsis are single-celled epidermal hair that are a useful model for studying plant cell fate determination. Trichome initiation requires the activity of the GLABROUS1 (GL1) gene whose expression in epidermal and trichome cells is dependent on the presence of a 3'-cis-regulatory element. Using a one-hybrid screen, we have isolated a cDNA, which encodes for a protein, GL1 enhancer binding protein (GeBP), that binds this regulatory element in yeast and in vitro. GeBP and its three homologues in Arabidopsis share two regions: a central region with no known motifs and a C-terminal region with a putative leucine-zipper motif. We show that both regions are necessary for trans-activation in yeast. A translational fusion with the Yellow Fluorescent Protein (YFP) indicates that GeBP is a nuclear protein whose localization is restricted to, on average, 3-5 subnuclear foci that might correspond to nucleoli. Transcriptional fusion with the GUS reporter indicates that GeBP is mainly expressed in vegetative meristematic tissues and in very young leaf primordia. We looked at GeBP expression in plants mutated in or misexpressing KNAT1, a KNOX gene, expressed in the shoot apical meristem and downregulated in leaf founder cells, and found that GeBP transcript level is regulated by KNAT1 suggesting that KNAT1 is a transcriptional activator of GeBP. This regulation suggests that GeBP is acting as a repressor of leaf cell fate.
拟南芥的毛状体是单细胞表皮毛,是研究植物细胞命运决定的有用模型。毛状体起始需要GLABROUS1(GL1)基因的活性,其在表皮和毛状体细胞中的表达依赖于一个3' - 顺式调控元件的存在。通过单杂交筛选,我们分离出一个cDNA,它编码一种蛋白质,即GL1增强子结合蛋白(GeBP),该蛋白在酵母和体外能结合这个调控元件。GeBP及其在拟南芥中的三个同源物共有两个区域:一个没有已知基序的中央区域和一个具有假定亮氨酸拉链基序的C端区域。我们表明这两个区域对于酵母中的反式激活都是必需的。与黄色荧光蛋白(YFP)的翻译融合表明GeBP是一种核蛋白,其定位平均局限于3 - 5个可能对应于核仁的亚核焦点。与GUS报告基因的转录融合表明GeBP主要在营养分生组织和非常幼嫩的叶原基中表达。我们研究了在KNAT1(一种KNOX基因,在茎尖分生组织中表达且在叶原基细胞中下调)发生突变或错误表达的植物中GeBP的表达,发现GeBP转录水平受KNAT1调控,这表明KNAT1是GeBP的转录激活因子。这种调控表明GeBP作为叶细胞命运的抑制因子发挥作用。