Graduate School of Bioagricultural Sciences, Nagoya University, Furo-Cho, Chikusa-Ku, Nagoya, Aichi, 464-8601, Japan.
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, Higashi 1-1-1, Tsukuba, Ibaraki, 305-8566, Japan.
Plant Mol Biol. 2024 May 7;114(3):54. doi: 10.1007/s11103-024-01450-z.
Sugars, synthesized by photosynthesis in source organs, are loaded and utilized as an energy source and carbon skeleton in sink organs, and also known to be important signal molecules regulating gene expression in higher plants. The expression of genes coding for sporamin and β-amylase, the two most abundant proteins in storage roots of sweet potato, is coordinately induced by sugars. We previously reported on the identification of the carbohydrate metabolic signal-responsible element-1 (CMSRE-1) essential for the sugar-responsible expression of two genes. However, transcription factors that bind to this sequence have not been identified. In this study, we performed yeast one-hybrid screening using the sugar-responsible minimal promoter region of the ß-amylase gene as bait and a library composed only transcription factor cDNAs of Arabidopsis. Two clones, named Activator protein binding to CMSRE-1 (ACRE), encoding AP2/ERF transcription factors were isolated. ACRE showed transactivation activity of the sugar-responsible minimal promoter in a CMSRE-1-dependent manner in Arabidopsis protoplasts. Electric mobility shift assay (EMSA) using recombinant proteins and transient co-expression assay in Arabidopsis protoplasts revealed that ACRE could actually act to the CMSRE-1. Among the DEHYDRATION -RESPONSIVE ELEMENT BINDING FACTOR (DREB) subfamily, almost all homologs including ACRE, could act on the DRE, while only three ACREs could act to the CMSRE-1. Moreover, ACRE-homologs of Japanese morning glory also have the same property of DNA-binding preference and transactivation activity through the CMSRE-1. These findings suggested that ACRE plays an important role in the mechanism regulating the sugar-responsible gene expression through the CMSRE-1 conserved across plant species.
糖是在源器官中通过光合作用合成的,被装载并在汇器官中用作能量源和碳骨架,也被认为是调节高等植物基因表达的重要信号分子。编码甘薯贮藏根中两种最丰富蛋白质的蔗糖酶和β-淀粉酶的基因表达受糖的协调诱导。我们之前报道了负责两个基因糖响应表达的碳水化合物代谢信号响应元件-1(CMSRE-1)的鉴定。然而,尚未鉴定与该序列结合的转录因子。在这项研究中,我们使用β-淀粉酶基因的糖响应最小启动子区域作为诱饵,使用仅包含拟南芥转录因子 cDNA 的文库进行酵母单杂交筛选。分离出两个克隆,命名为与 CMSRE-1 结合的激活蛋白(ACRE),编码 AP2/ERF 转录因子。ACRE 在 CMSRE-1 依赖的方式下在拟南芥原生质体中表现出糖响应最小启动子的转录激活活性。使用重组蛋白进行的电泳迁移率变动分析(EMSA)和在拟南芥原生质体中的瞬时共表达测定表明,ACRE 实际上可以作用于 CMSRE-1。在脱水响应元件结合因子(DREB)亚家族中,几乎所有包括 ACRE 在内的同源物都可以作用于 DRE,而只有三个 ACRE 可以作用于 CMSRE-1。此外,日本牵牛的 ACRE 同源物也具有相同的 DNA 结合偏好和通过 CMSRE-1 的转录激活活性的特性。这些发现表明,ACRE 通过在植物物种中保守的 CMSRE-1 发挥重要作用,调节糖响应基因表达的机制。