Ito M, Iwase M, Kodama H, Lavisse P, Komamine A, Nishihama R, Machida Y, Watanabe A
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Tokyo 113, Japan.
Plant Cell. 1998 Mar;10(3):331-41. doi: 10.1105/tpc.10.3.331.
Plant B-type cyclin genes are expressed late in the G2 and M phases of the cell cycle. Previously, we showed that the promoter of a Catharanthus roseus B-type cyclin, CYM, could direct M phase-specific transcription of a beta-glucuronidase reporter gene in synchronously dividing BY2 tobacco cells. In this study, we determined the regulatory elements contained within the CYM promoter by using a luciferase reporter gene. Mutational analysis showed that a 9-bp element is essential for M phase-specific promoter activity in synchronized BY2 cells. The CYM promoter contains three other sequences similar to this element. A gain-of-function assay demonstrated that when fused to a heterologous promoter, these elements are sufficient for M phase-specific expression; therefore, we named these elements M-specific activators (MSAs). We found MSA-like sequences in B-type cyclin promoters from tobacco, soybean, and Arabidopsis as well as in the promoters of two M phase-specific genes, NACK1 and NACK2, which encode tobacco kinesin-like proteins. Thus, MSA may be a common cis-acting promoter element that controls M phase-specific expression of cell cycle-related genes in plants.
植物B型细胞周期蛋白基因在细胞周期的G2期和M期晚期表达。此前,我们发现长春花B型细胞周期蛋白CYM的启动子能够在同步分裂的BY2烟草细胞中指导β-葡萄糖醛酸酶报告基因的M期特异性转录。在本研究中,我们利用荧光素酶报告基因确定了CYM启动子中所含的调控元件。突变分析表明,一个9碱基对元件对于同步化的BY2细胞中M期特异性启动子活性至关重要。CYM启动子还包含另外三个与该元件相似的序列。功能获得分析表明,当与异源启动子融合时,这些元件足以实现M期特异性表达;因此,我们将这些元件命名为M期特异性激活因子(MSA)。我们在烟草、大豆和拟南芥的B型细胞周期蛋白启动子以及两个M期特异性基因NACK1和NACK2的启动子中发现了类似MSA的序列,NACK1和NACK2编码烟草类驱动蛋白。因此,MSA可能是一种常见的顺式作用启动子元件,可控制植物中细胞周期相关基因的M期特异性表达。